LOCUS       V027336                 9201 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027336
VERSION     V027336
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2301..4055
                     /label="FAM83D(NM_030919)"
                     /note="FAM83D(NM_030919)"
                     /gene="FAM83D"
     CDS             4062..4127
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4171..4382
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4395..4663
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4690..5289
                     /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    5296..5884
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5958..6191
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6263..6384
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6403..6538
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6576..6592)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6600..6616
                     /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(6624..6654)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6669..6690
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6978..7566)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7737..8597)
                     /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(8598..8702)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9176..9192
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atggctctgc tgtccgaggg cctggacgag gtgcccgccg
     2341 cctgcctgtc gccgtgcggg ccgcccaacc cgaccgagct gttcagcgag tcacggcgcc
     2401 tggctctgga ggagctggtg gcgggcggcc ccgaagcctt cgcggccttc ctgcgacgcg
     2461 agcgcctggc tcgtttcctg aaccccgatg aggtgcacgc cattctgcgc gcggcggaga
     2521 ggccgggaga ggagggcgcg gcggcggcgg cggcggccga ggactcgttc ggctcctcgc
     2581 acgactgctc ttcgggcacc tacttccccg agcagtcgga cctggagcca ccgctgttgg
     2641 agcttggctg gcccgccttc taccagggcg cctaccgcgg cgccacgcgt gtcgagacgc
     2701 acttccagcc ccgcggcgct ggcgaaggtg gcccctacgg ctgcaaggac gctctgcgcc
     2761 agcagctccg ctcggcgcga gaggtgattg cagtggtcat ggacgtgttc acagacatcg
     2821 acatcttcag agacctgcaa gaaatatgca ggaaacaggg agttgctgtg tatatccttc
     2881 tggaccaggc tctcctctct caatttctgg atatgtgcat ggatctgaaa gttcatcctg
     2941 aacaggaaaa gttaatgaca gttcggacta tcacaggaaa tatctactat gcaaggtcag
     3001 gaactaagat tattgggaag gttcacgaaa agttcacgtt gattgatggc atccgcgtgg
     3061 caacaggctc ctacagtttt acatggacgg atggcaaatt aaacagcagt aacttggtaa
     3121 ttctgtctgg ccaagtggtt gaacactttg atctggagtt ccgaatcctg tatgcccagt
     3181 ccaagcccat cagccccaaa ctcctgtctc acttccagag cagcaacaag tttgatcacc
     3241 tcaccaaccg aaaaccacag tccaaggagc tcaccctggg caacctgctg cggatgcggc
     3301 tggctaggct gtcaagtact cccaggaagg cggacctgga cccagagatg cccgcagagg
     3361 gcaaggcaga gcgcaagccc catgactgtg agtcctctac tgttagtgag gaagactact
     3421 tcagcagcca cagggacgag ctccagagca gaaaggccat tgacgctgcc actcaaacag
     3481 agccaggaga ggagatgcca gggctgagtg tgagtgaggt gggaacacaa accagcatca
     3541 ccacagcatg tgctggtacc cagactgcag tcatcaccag gatagcaagc tctcaaacca
     3601 cgatttggtc cagatcgacc actactcaga ctgacatgga tgagaacatt ctctttcctc
     3661 gaggaactca atctacagaa gggtcaccag tctcaaaaat gtctgtatcg agatcttcca
     3721 gtttgaagtc ttcctcctct gtgtcttccc aaggctctgt ggcaagctcc actggttctc
     3781 ccgcttccat cagaaccact gacttccaca atcctggcta tcccaagtac ctgggcaccc
     3841 cccacctgga actgtacttg agtgactcac ttagaaactt gaacaaagag cggcaattcc
     3901 acttcgctgg tatcaggtcc cggctcaacc acatgctggc tatgctgtca aggagaacac
     3961 tctttactga aaaccacctt ggccttcatt ctggcaattt cagcagagtt aatttgcttg
     4021 ctgttagaga tgtagcactt tatccttcct atcagggatc cgactacaaa gaccatgacg
     4081 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag cggccgcgaa
     4141 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     4201 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
     4261 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     4321 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     4381 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
     4441 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
     4501 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
     4561 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
     4621 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc
     4681 ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4741 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4801 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4861 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4921 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     4981 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5041 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5101 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5161 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5221 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5281 ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     5341 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     5401 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5461 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5521 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5581 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5641 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5701 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5761 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5821 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5881 ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     5941 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt
     6001 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     6061 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     6121 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6181 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     6241 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     6301 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6361 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     6421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6481 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6541 gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6601 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6661 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6721 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6781 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6841 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6901 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6961 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7021 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7081 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7141 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7201 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7321 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7381 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7441 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7501 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7561 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7621 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7681 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7741 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7801 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7861 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7921 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7981 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8041 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8101 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8161 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8221 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8281 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8341 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8401 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8461 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8521 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8581 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8641 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8701 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     8761 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     8821 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     8881 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     8941 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     9001 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     9061 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     9121 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     9181 acgacggcca gtgccaagct g
//
