LOCUS       V037324                 9607 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037324
VERSION     V037324
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             2302..4461
                     /label="TGM5(NM_201631)"
                     /note="TGM5(NM_201631)"
                     /gene="TGM5"
     CDS             4468..4533
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4577..4788
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4801..5069
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5096..5695
                     /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    5702..6290
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6364..6597
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6669..6790
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6809..6944
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6982..6998)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7006..7022
                     /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(7030..7060)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7075..7096
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7384..7972)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8143..9003)
                     /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(9004..9108)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9582..9598
                     /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggcccaa gggctagaag tggccctcac agacctccag
     2341 agctccagaa ataatgtgcg gcaccacacg gaggagatca ctgtggacca cctgcttgtt
     2401 cgccggggcc aggccttcaa cctcaccctg tacttcagga accggagctt ccagccaggc
     2461 ctggacaaca tcatcttcgt ggttgaaact ggaccgctgc cagacctggc cttggggact
     2521 cgggctgtgt tcagcctggc acgccatcac agccccagcc cctggattgc ctggctggag
     2581 accaatgggg ccacctccac agaggtgagc ttgtgcgctc ctcccacggc ggccgtgggt
     2641 cggtacctct tgaaaatcca catcgactcc ttccaggggt ctgtgacggc ctaccagcta
     2701 ggggagttca tcctgctttt caatccctgg tgcccagagg atgctgtcta cttggacagt
     2761 gaaccccaga ggcaggagta tgtcatgaat gattatggct tcatctacca aggcagcaag
     2821 aactggatcc gcccatgtcc ctggaactat ggacagtttg aagacaaaat catagacatc
     2881 tgcctgaagc tgctagacaa gagcctgcac ttccagactg acccagccac agactgtgct
     2941 ctgcggggaa gccccgtcta cgtcagcaga gtggtgtgtg ccatgatcaa cagcaatgat
     3001 gataatgggg tgctcaatgg aaactggagt gagaattaca cagacggcgc caaccctgcg
     3061 gagtggacgg gcagcgtggc catcctgaag cagtggaacg ccacaggctg ccagcccgtg
     3121 cgctacgggc aatgctgggt ctttgctgcc gtcatgtgca cagtgatgag gtgtctgggg
     3181 atccctaccc gtgtgatcac caacttcgac tctggccacg atacagatgg aaacctgatc
     3241 atagatgagt attatgacaa cacaggcagg attttgggga ataagaagaa ggatactatc
     3301 tggaacttcc atgtctggaa tgagtgctgg atggcccgga aggatctgcc ccctgcatat
     3361 ggaggctggc aggtgctgga cgccacacct caggagatga gcaacggcgt ctactgctgt
     3421 ggccctgcct ctgtcagagc catcaaagaa ggagaagtgg acctgaacta tgacacgccc
     3481 tttgtgtttt cgatggtgaa tgctgactgc atgtcctggc tcgtccaggg agggaaggag
     3541 cagaagcttc accaggacac gagttctgtt ggcaatttta tcagcacaaa gagcatccag
     3601 agtgacgagc gggatgacat cacagagaac tacaagtatg aagaaggatc cctccaggag
     3661 aggcaggtgt ttctgaaggc tctgcagaag ctgaaggcta gaagcttcca tggctcccaa
     3721 agaggagcag agttgcaacc ttccaggccc acatcactga gccaggacag ccctcggagc
     3781 ctgcatacac cttcccttcg acccagtgat gtggtgcaag tctccctgaa attcaagctg
     3841 ctcgacccgc ccaacatggg ccaggatata tgctttgtcc tgctggccct caacatgtcc
     3901 tcccagttca aggacctcaa agtgaacctg agtgcccagt ctctgctgca cgatggcagc
     3961 cccctgtccc cattctggca ggacacagcg ttcatcacac tctctcctaa agaagcaaag
     4021 acctacccct gcaaaatctc ctattcccag tacagccagt acctgtcaac agacaagctg
     4081 atccgcatca gtgccctggg tgaagagaaa agcagtcctg agaaaatcct ggtgaacaag
     4141 atcatcacct tatcttatcc aagcatcacg attaatgttc taggagcagc cgttgtgaac
     4201 cagccactct ccatacaggt gatattttca aaccccctct cggagcaggt tgaggactgt
     4261 gtgctgactg tggaaggaag tggcctcttc aagaaacagc agaaagtctt ccttggagtc
     4321 ctcaaacccc aacaccaagc aagcatcatt ctggagaccg tccccttcaa gagtggacaa
     4381 aggcagatcc aagctaatat gagaagcaac aagtttaagg acattaaggg ttacaggaat
     4441 gtttatgtag actttgcatt aggatccgac tacaaagacc atgacggtga ttataaagat
     4501 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc
     4561 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     4621 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     4681 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     4741 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     4801 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     4861 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     4921 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     4981 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     5041 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac
     5101 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac
     5161 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca
     5221 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     5281 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     5341 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     5401 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     5461 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     5521 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5581 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5641 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga
     5701 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5761 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5821 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5881 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5941 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6001 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6061 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     6121 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6181 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt
     6301 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6361 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
     6421 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6481 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6541 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6601 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6661 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6721 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6781 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
     6841 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6901 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     6961 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     7021 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7081 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     7141 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     7201 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7261 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7321 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7381 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7441 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7501 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7561 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7621 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7681 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7741 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7801 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7861 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7921 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7981 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     8041 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     8101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     8161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8281 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     9001 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     9061 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     9121 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     9181 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     9241 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     9301 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9361 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     9421 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     9481 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     9541 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9601 caagctg
//
