LOCUS       V033119                 8915 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033119
VERSION     V033119
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        complement(join(8829..8915,1..18))
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     492..508
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     LTR             751..931
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    975..1100
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1593..1826
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             2010..2054
                     /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    2315..2432
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2522..2725
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2799..2843
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
     CDS             2877..2900
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2904..2927
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2937..3050
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     CDS             3132..4301
                     /label="Tyr(NM_001317397)"
                     /note="Tyr(NM_001317397)"
                     /gene="Tyr"
     CDS             4308..4331
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             4335..4358
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4402..4613
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4626..4894
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4921..5520
                     /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    5527..6115
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6189..6422
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6494..6615
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6634..6769
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6807..6823)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6831..6847
                     /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(6855..6885)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6900..6921
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7209..7797)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7968..8828)
                     /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"
ORIGIN
        1 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
       61 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
      121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
      181 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      241 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
      361 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
      421 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
      481 gtcacgacgt tgtaaaacga cggccagtgc caagctgacg cgtgtagtct tatgcaatac
      541 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
      601 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
      661 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
      721 tatttaagtg cctagctcga tacaataaac gggtctctct ggttagacca gatctgagcc
      781 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
      841 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
      901 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ctgaaagcga
      961 aagggaaacc agagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1021 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1081 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1141 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1201 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1261 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1321 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1381 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1441 ccactgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     1501 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     1561 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     1621 gcagcaggaa gcactatggg cgcagcctca atgacgctga cggtacaggc cagacaatta
     1681 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     1741 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     1801 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     1861 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gattggaatc
     1921 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1981 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2041 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2101 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2161 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2221 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2281 ccattcgatt agtgaacgga tctcgacggt taacttttaa aagaaaaggg gggattgggg
     2341 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2401 tacaaaaaca aattacaaaa attcaaaatt ttatcgatac tagtattatg cccagtacat
     2461 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2521 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     2581 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     2641 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     2701 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     2761 tccacgctgt tttgacctcc atagaagatt ctagaatgaa agaaaccgct gctgctaaat
     2821 tcgaacgcca gcacatggac agcggagccg gcgcaggagc cggcgctgac gcgcctgact
     2881 acaaagacga tgacgacaag ggagattaca aggatgacga tgacaaaggc gcgtcgatgg
     2941 acgagaagac caccggctgg cggggcggcc acgtggtgga gggcctggcc ggcgagctgg
     3001 agcagctgcg ggccaggctg gagcaccacc ctcagggcca gcgggagccc tccggcggct
     3061 gcaagctggg cggcgcgcct gaattctgca gtcgacggta ccgcgggccc atcgctcgat
     3121 ctagagctag catgttcttg gctgttttgt attgccttct gtggagtttc cagatctctg
     3181 atggccattt tcctcgagcc tgtgcctcct ctaagaactt gttggcaaaa gaatgctgcc
     3241 caccatggat gggtgatggg agtccctgcg gccagctttc aggcagaggt tcctgccagg
     3301 atatccttct gtccagtgca ccatctggac ctcagttccc cttcaaaggg gtggatgacc
     3361 gtgagtcctg gccctctgtg ttttataata ggacctgcca gtgctcaggc aacttcatgg
     3421 gtttcaactg cggaaactgt aagtttggat ttgggggccc aaattgtaca gagaagcgag
     3481 tcttgattag aagaaacatt tttgatttga gtgtctccga aaagaataag ttcttttctt
     3541 acctcacttt agcaaaacat actatcagct cagtctatgt catccccaca ggcacctatg
     3601 gccaaatgaa caatgggtca acacccatgt ttaatgatat caacatctac gacctctttg
     3661 tatggatgca ttactatgtg tcaagggaca cactgcttgg gggctctgaa atatggaggg
     3721 acattgattt tgcccatgaa gcaccagggt ttctgccttg gcacagactt ttcttgttat
     3781 tgtgggaaca agaaattcga gaactaactg gggatgagaa cttcactgtt ccatactggg
     3841 attggagaga tgcagaaaac tgtgacattt gcacagatga gtacttggga ggtcgtcacc
     3901 ctgaaaatcc taacttactc agcccagcat ccttcttctc ctcctggcag atcatttgta
     3961 gcagatcaga agagtataat agccatcagg ttttatgcga tggaacacct gagggaccac
     4021 tattacgtaa tcctggaaac catgacaaag ccaaaacccc caggctccca tcttcagcag
     4081 atgtggaatt ttgtctgagt ttgacccagt atgaatctgg atcaatggat agaactgcca
     4141 atttcagctt tagaaacaca ctggaagtat ttttgaacaa tggctgcgaa ggcaccgccc
     4201 tcttttggaa gtttacccag aagccaatgc acctatcggc cataacagag actcttacat
     4261 ggttcctttc ataccgctct atagaaatgg tgatttcttc aggatccgat tataaggatg
     4321 atgacgacaa aggagactat aaagacgacg atgacaagta gcggccgcga aggatctgcg
     4381 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     4441 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     4501 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     4561 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     4621 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     4681 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     4741 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     4801 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     4861 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag
     4921 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     4981 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     5041 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5101 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5161 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5221 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5281 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5341 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5401 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5461 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5521 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5581 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5641 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5701 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5761 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5821 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5881 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     5941 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     6001 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6061 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt
     6121 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6181 ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac
     6241 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6301 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6361 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6421 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     6481 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6541 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6601 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc
     6661 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6721 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     6781 agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7621 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8881 agcggataca tatttgaatg tatttagaaa aataa
//
