LOCUS       V037615                 9307 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037615
VERSION     V037615
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..4161
                     /label="SLC6A6(NM_003043)"
                     /note="SLC6A6(NM_003043)"
                     /gene="SLC6A6"
     CDS             4168..4233
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4277..4488
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4501..4769
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4796..5395
                     /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    5402..5990
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6064..6297
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6369..6490
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6509..6644
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6682..6698)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6706..6722
                     /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(6730..6760)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6775..6796
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7084..7672)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7843..8703)
                     /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(8704..8808)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9282..9298
                     /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 catggccacc aaggagaagc tgcagtgtct gaaagatttc
     2341 cacaaggaca tcctgaagcc ctcaccaggg aagagcccag gcacgcggcc tgaggacgag
     2401 gctgagggaa aacctccgca gagggagaag tggtctagca agatcgactt tgtgctctct
     2461 gtggctggcg gcttcgtggg cttgggcaac gtctggcgct tcccgtacct ctgctacaag
     2521 aatggtggag gtgcgtttct cataccgtat tttattttcc tgtttgggag cggcctgcct
     2581 gtgtttttct tggagatcat cataggccag tacacctctg aagggggcat cacctgctgg
     2641 gaaaagatct gccccttgtt ctctggtatc ggctatgcct ccgttgtaat tgtgtccctc
     2701 ctgaatgtct actacatcgt catcctggcc tgggccacat actacctgtt ccagtccttc
     2761 cagaaggagc tgccctgggc acactgcaac cacagctgga acacacctca ctgcatggag
     2821 gacaccatgc gcaagaacaa gagtgtctgg atcaccatca gctccaccaa cttcacctcc
     2881 cctgtcatcg agttctggga gcgcaacgtg ctgagcttgt cccctggaat cgaccaccca
     2941 ggctctctga aatgggacct cgctctctgc cttcttttag tctggctagt gtgtttcttc
     3001 tgcatctgga agggcgtcag gtccactggg aaggtcgtct acttcacagc cacttttcca
     3061 ttcgccatgc tcctggtgct gctggtccga gggctgacgc tgccgggcgc gggcgcaggc
     3121 atcaagttct atctgtatcc tgacatcacc cgccttgagg acccacaggt gtggattgac
     3181 gctgggactc agatattctt ctcttatgcc atctgcctgg gggctatgac ctcgctgggg
     3241 agctacaaca agtacaagta taactcgtac agggactgta tgctgctggg atgcctgaac
     3301 agtggtacca gttttgtgtc tggcttcgca attttttcca tcctgggctt catggcacaa
     3361 gagcaagggg tggacattgc tgatgtggct gagtcaggtc ctggcctggc cttcattgcc
     3421 tacccaaaag ctgtgacaat gatgccgctg cccacatttt ggtccattct tttttttatt
     3481 atgcttctct tgcttggact ggatagccag tttgttgaag ttgaaggaca gatcacatcc
     3541 ttggttgatc tttacccatc cttcctaagg aagggttatc gtcgggaaat cttcatcgcc
     3601 ttcgtgtgta gcatcagcta cctgctgggg ctgacgatgg tgacggaggg tggcatgtat
     3661 gtgtttcagc tctttgacta ctatgcagct agcggtgtat gccttttgtg ggttgcattc
     3721 tttgaatgtt ttgttattgc ctggatatat ggaggtgata acctttatga tggtattgag
     3781 gacatgattg gctatcggcc cgggccctgg atgaagtaca gctgggctgt gatcactcca
     3841 gttctctgtg ttggatgttt catcttctcg ctcgtcaagt acgtacccct gacctacaac
     3901 aaaacatacg tgtaccccaa ctgggccatt gggctgggct ggagcctggc cctttcctcc
     3961 atgctctgcg ttcccttggt catcgtcatc cgcctctgcc agactgaggg gccgttcctt
     4021 gtgagagtca agtacctgct gaccccaagg gaacccaacc gctgggctgt ggagcgcgag
     4081 ggagccacac cttacaactc tcgcaccgtc atgaacggcg ctctcgtgaa accgacccac
     4141 atcattgtgg agaccatgat gggatccgac tacaaagacc atgacggtga ttataaagat
     4201 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc
     4261 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     4321 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     4381 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     4441 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     4501 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     4561 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     4621 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     4681 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     4741 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac
     4801 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac
     4861 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca
     4921 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4981 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     5041 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     5101 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     5161 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     5221 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5281 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5341 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga
     5401 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5461 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5521 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5581 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5641 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5701 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5761 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5821 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5881 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5941 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt
     6001 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6061 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
     6121 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6181 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6241 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6301 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6361 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6421 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6481 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
     6541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     6661 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6721 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6781 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6841 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6901 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7261 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7501 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7621 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7681 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7741 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7801 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7861 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7921 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7981 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8041 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8101 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8161 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8221 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8281 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8341 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8401 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8461 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8521 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8581 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8641 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8701 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8761 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8821 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8881 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8941 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     9001 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9061 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     9121 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     9181 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     9241 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9301 caagctg
//
