LOCUS       V024803                 9452 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024803
VERSION     V024803
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..3495
                     /label="KCNJ9(NM_004983)"
                     /note="KCNJ9(NM_004983)"
                     /gene="KCNJ9"
     CDS             3502..3567
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3611..3822
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3835..4103
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4132..4141
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4162..4824
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4894..4947
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4948..5547
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5548..6135
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6209..6442
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6514..6635
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6654..6789
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6827..6843)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6851..6867
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6875..6905)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6920..6941
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7229..7817)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7988..8848)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8849..8953)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9427..9443
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg cgcaggagaa cgcggccttc
     2341 tcgcccgggc aggaggagcc gccgcggcgc cgcggccgcc agcgctacgt ggagaaggat
     2401 ggccggtgca acgtgcagca gggcaacgtg cgcgagacat accgctacct gacggacctg
     2461 ttcaccacgc tggtggacct gcagtggcgc ctcagcctgt tgttcttcgt cctggcctac
     2521 gcgctcacct ggctcttctt cggcgccatc tggtggctga tcgcctacgg ccgcggcgac
     2581 ctggagcacc tggaggacac cgcgtggacg ccgtgcgtca acaacctcaa cggcttcgtg
     2641 gccgccttcc tcttctccat cgagaccgag accaccatcg gctacgggca ccgcgtcatc
     2701 accgaccagt gccccgaggg catcgtgctg ctgctgctgc aggccatcct gggctccatg
     2761 gtgaacgcct tcatggtggg ctgcatgttc gtcaagatct cgcagcccaa caagcgcgca
     2821 gccacgctcg tcttctcctc gcacgccgtg gtgtcgctgc gcgacgggcg cctctgcctc
     2881 atgttccgcg tgggcgactt gcgctcctca cacatagtgg aggcctccat ccgcgccaag
     2941 ctcatccgct cgcgccagac gctggagggc gagttcatcc cgctgcacca gaccgacctc
     3001 agcgtgggct tcgacacggg agacgaccgc ctcttcctcg tctcgccgct ggttatcagc
     3061 cacgagatcg acgccgccag ccccttctgg gaggcgtcgc gccgtgccct cgagagggac
     3121 gacttcgaga tcgtcgttat cctcgagggc atggtggaag ccacgggaat gacatgccaa
     3181 gctcggagct cctacctggt agacgaggtg ctgtggggcc accgcttcac gtcagtgctg
     3241 actctggagg acggcttcta cgaagtggac tatgccagct ttcacgagac ttttgaggtg
     3301 cccacacctt cgtgcagtgc tcgagagctg gcagaggctg ccgcccgcct tgatgcccat
     3361 ctctactggt ccatccccag ccggctggat gagaaggtgg aggaggaggg ggcgggggag
     3421 ggggcgggtg gggaagctgg ggctgacaag gagcagaatg gctgcctgcc acccccagag
     3481 agtgagtcca aggtgggatc cgactacaaa gaccatgacg gtgattataa agatcatgac
     3541 atcgactaca aggatgacga tgacaagtag cggccgcgaa ggatctgcga tcgctccggt
     3601 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     3661 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     3721 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     3781 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     3841 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     3901 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     3961 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     4021 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     4081 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga cgccaccatg
     4141 gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac cggcaccctg
     4201 aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccaagca gggccgcatg
     4261 accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct gctgagccac
     4321 gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga gaaccccttc
     4381 ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta cgaggacggc
     4441 ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat cggcgacttc
     4501 aaggtggtgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa gatcatccgc
     4561 agcaacgcca ccgtggagca cctgcacccc atgggcgata acgtgctggt gggcagcttc
     4621 gcccgcacct tcagcctgcg cgacggcggc tactacagct tcgtggtgga cagccacatg
     4681 cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat gttcgccttc
     4741 cgccgcgtgg aggagctgca cagcaacacc gagctgggca tcgtggagta ccagcacgcc
     4801 ttcaagaccc ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc tgccgtggac
     4861 ggcaccgccg gacccggctc caccggatct cgcgagggca gaggaagtct tctaacatgc
     4921 ggtgacgtgg aggagaatcc cggccctatg accgagtaca agcccacggt gcgcctcgcc
     4981 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5041 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5101 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     5161 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     5221 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     5281 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     5341 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     5401 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     5461 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     5521 tgcatgaccc gcaagcccgg tgcctgaaat caacctctgg attacaaaat ttgtgaaaga
     5581 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5641 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5701 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5761 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5821 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5881 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     5941 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6001 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6061 ccggctctgc ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg gatctccctt
     6121 tggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     6181 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa
     6241 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6301 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6361 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6421 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     6481 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     6541 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6601 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     6661 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6721 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6781 ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc
     6841 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6901 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6961 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7021 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7081 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7141 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7201 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     9001 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     9061 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     9121 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     9181 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     9241 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     9301 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     9361 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     9421 gacgttgtaa aacgacggcc agtgccaagc tg
//
