LOCUS       V032634                 8057 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032634
VERSION     V032634
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             2287..2352
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2377..2982
                     /label="C8G(NM_000606)"
                     /note="C8G(NM_000606)"
                     /gene="C8G"
     promoter        3027..3238
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3251..3519
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3546..4145
                     /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    4152..4740
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             4814..5047
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5119..5240
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5259..5394
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5432..5448)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5456..5472
                     /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(5480..5510)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5525..5546
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5834..6422)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6593..7453)
                     /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(7454..7558)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8032..8048
                     /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 agaatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     2341 gacgatgaca agtctagaga attcggatcc gccaccatgc tgccccctgg gactgcgacc
     2401 ctcttgactc tgctcctggc agctggctcg ctgggccaga agcctcagag gccacgccgg
     2461 cccgcatccc ccatcagcac catccagccc aaggccaatt ttgatgctca gcagtttgca
     2521 gggacctggc tccttgtggc tgtgggctcc gcttgccgtt tcctgcagga gcagggccac
     2581 cgggccgagg ccaccacact gcatgtggct ccccagggca cagccatggc tgtcagtacc
     2641 ttccgaaagc tggatgggat ctgctggcag gtgcgccagc tctatggaga cacaggggtc
     2701 ctcggccgct tcctgcttca agcccgagac gcccgagggg ctgtgcacgt ggttgtcgct
     2761 gagaccgact accagagttt cgctgtcctg tacctggagc gggcggggca gctgtcagtg
     2821 aagctctacg cccgctcgct ccctgtgagc gactcggtcc tgagtgggtt tgagcagcgg
     2881 gtccaggagg cccacctgac tgaggaccag atcttctact tccccaagta cggcttctgc
     2941 gaggctgcag accagttcca cgtcctggac gaagtgagga ggtaagcggc cgcgaaggat
     3001 ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     3061 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg
     3121 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     3181 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct
     3241 gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca
     3301 tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc
     3361 cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg
     3421 gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta
     3481 cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac
     3541 gctagatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     3601 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     3661 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     3721 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     3781 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     3841 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     3901 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     3961 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     4021 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     4081 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     4141 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4201 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4261 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4321 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4381 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4441 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4501 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     4561 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4621 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4681 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4741 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     4801 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct
     4861 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     4921 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     4981 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5041 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     5101 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5161 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5221 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     5281 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5341 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     5401 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5461 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5521 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5581 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6241 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7561 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     7621 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     7681 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     7741 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     7801 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     7861 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     7921 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     7981 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     8041 cggccagtgc caagctg
//
