LOCUS       V038132                 8807 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038132
VERSION     V038132
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"
     regulatory      2288..2297
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2297..2362
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2369..3709
                     /label="SLC16A11(NM_001370549)"
                     /note="SLC16A11(NM_001370549)"
                     /gene="SLC16A11"
     promoter        3777..3988
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4001..4269
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4296..4895
                     /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    4902..5490
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5564..5797
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5869..5990
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6009..6144
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6182..6198)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6206..6222
                     /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(6230..6260)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6275..6296
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6584..7172)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7343..8203)
                     /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(8204..8308)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8782..8798
                     /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2341 ctacaaggat gacgatgaca aggaattcat gaccccccag cccgccggac ccccggatgg
     2401 gggctggggc tgggtggtgg cggccgcagc cttcgcgata aacgggctgt cctacgggct
     2461 gctgcgctcg ctgggccttg ccttccctga ccttgccgag cactttgacc gaagcgccca
     2521 ggacactgcg tggatcagcg ccctggccct ggccgtgcag caggcagcca gccccgtggg
     2581 cagcgccctg agcacgcgct ggggggcccg ccccgtggtg atggttgggg gcgtcctcgc
     2641 ctcgctgggc ttcgtcttct cggctttcgc cagcgatctg ctgcatctct acctcggcct
     2701 gggcctcctc gctggctttg gttgggccct ggtgttcgcc cccgccctag gcaccctctc
     2761 gcgttacttc tcccgccgtc gagtcttggc ggtggggctg gcgctcaccg gcaacggggc
     2821 ctcctcgctg ctcctggcgc ccgccttgca gcttcttctc gatactttcg gctggcgggg
     2881 cgctctgctc ctcctcggcg cgatcaccct ccacctcacc ccctgtggcg ccctgctgct
     2941 acccctggtc cttcctggag accccccagc cccaccgcgt agtcccctag ctgccctcgg
     3001 cctgagtctg ttcacacgcc gggccttctc aatctttgct ctaggcacag ccctggttgg
     3061 gggcgggtac ttcgttcctt acgtgcactt ggctccccac gctttagacc ggggcctggg
     3121 gggatacgga gcagcgctgg tggtggccgt ggctgcgatg ggggatgcgg gcgcccggct
     3181 ggtctgcggg tggctggcag accaaggctg ggtgcccctc ccgcggctgc tggccgtatt
     3241 cggggctctg actgggctgg ggctgtgggt ggtggggctg gtgcccgtgg tgggcggcga
     3301 agagagctgg gggggtcccc tgctggccgc ggctgtggcc tatgggctga gcgcggggag
     3361 ttacgccccg ctggttttcg gtgtactccc cgggctggtg ggcgtcggag gtgtggtgca
     3421 ggccacaggg ctggtgatga tgctgatgag cctcgggggg ctcctgggcc ctcccctgtc
     3481 aggcttccta agggatgaga caggagactt caccgcctct ttcctcctgt ctggttcttt
     3541 gatcctctcc ggcagcttca tctacatagg gttgcccagg gcgctgccct cctgtggtcc
     3601 agcctcccct ccagccacgc ctcccccaga gacgggggag ctgcttcccg ctccccaggc
     3661 agtcttgctg tccccaggag gccctggctc cactctggac accacttgtt gaggatccct
     3721 cgagagatca gtgtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3781 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3841 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     3901 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     3961 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     4021 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     4081 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     4141 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     4201 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     4261 ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc
     4321 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     4381 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     4441 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4501 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4561 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4621 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4681 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4741 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4801 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4861 gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca
     4921 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     4981 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5041 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5101 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5161 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5221 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5281 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     5341 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5401 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5461 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     5521 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5581 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5641 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5701 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5761 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     5821 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     5881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6001 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6061 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6121 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     6181 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6241 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6301 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6361 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7021 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7261 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7321 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7381 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7441 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7501 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7561 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7621 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7681 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7741 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7801 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7861 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7921 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7981 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8041 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8101 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8161 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8221 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8281 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8341 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8401 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     8461 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     8521 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     8581 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     8641 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     8701 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8761 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
