LOCUS       V037936                 9101 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037936
VERSION     V037936
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..4003
                     /label="Slc16a2(NM_009197)"
                     /note="Slc16a2(NM_009197)"
                     /gene="Slc16a2"
     promoter        4071..4282
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4295..4563
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4590..5189
                     /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    5196..5784
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5858..6091
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6163..6284
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6303..6438
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6476..6492)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6500..6516
                     /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(6524..6554)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6569..6590
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6878..7466)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7637..8497)
                     /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(8498..8602)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9076..9092
                     /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 ggcgctgcca agcccggcga gcgaggaagc
     2401 tgaggggccc tgtcaggagg caaaccagga atatcaggag ccggtgtgta gtcctgtgcc
     2461 cgagccggag ccagagccgg agcccgaacc ggagcctgac cccgagccgg tgccagtgcc
     2521 cccgcccgag cctcagcctg agcccgagcc ccagccccta cccgatcctg cacccctgcc
     2581 ggagctgggc tttgaggctg agccagtgca agaacccgag cccacgccca ctgtggaaac
     2641 ccgaggcacc gcgcgtggct ttcagcctcc cgaaggtggc ttcggctgga tagtggtgtt
     2701 tgcagccacc tggtgcaacg gctccatctt cggcatccat aactctgttg gtatcctcta
     2761 ctccatgcta ctagaggagg aaaaagagaa aaatcgccaa gtggagttcc aagcagcatg
     2821 ggtcggagcc ctcgctatgg gcatgatctt cttctgttct cccattgtga gtatattcac
     2881 tgaccgtttg ggctgccgaa tcacagccac caccggggcc gctgttgctt tcattggcct
     2941 ccacaccagc tccttcacca gctccctaag cctgcgctac ttcacctatg ggattctgtt
     3001 tggttgtggc tgctcattcg cctttcaacc atcactcgtc atcctgggcc actactttca
     3061 acgtcgccta ggtctagcca atggtgtggt atctgctgga agtagcatct tctccatgtc
     3121 ttttcccttc ctcatcaaaa tgctggggga taaaatcaag ctggcccaaa ccttccaggt
     3181 gctgagtacc ttcatgtttg tccttacgct gctctcactc acttaccggc ccctcctgcc
     3241 aagctcccag gacaccccaa gcaagagagg tgcccacacc ctgcgacagc gctttctggt
     3301 tcagttcaga aagtacttca acatgcgtgt attccgccag cgcacttacc gtatctgggc
     3361 ctttgggatc gctgctgctg cccttggtta cttcgtcccc tacgtgcacc tgatgaaata
     3421 tgtggaagat aagttcaagg agatcaagga gacctgggtg ctcttggtgt gcattggggc
     3481 tacgtcaggc ctcggacgtc tcgtctcggg ccacatcagt gactccatcc ctggacttaa
     3541 gaagatatac ttgcaggtcc tctcattcct gctcctgggc ttgatgtcca tgatgatccc
     3601 cctatgccgg gacttcgggg gccttatcgt tgtctgcctc ttcctgggcc tatgtgatgg
     3661 cttcttcatt accatcatgg ctcccattgc atttgagctg gtgggaccca tgcaggcctc
     3721 acaggccatt ggctacctcc tgggcatgat ggccctgcca atgattgctg ggccccccat
     3781 tgcaggcctc cttcgaaact gctttggtga ttaccatgtg gccttctact ttgcgggtgt
     3841 ccccccaatc attggagctg tgatcctttt cttcgtccct ctgatgcatc aaaggatgtt
     3901 taagaaagag cagagagatt ccagcaagga taagatgctg tcccacgatc cagaccccaa
     3961 cggagaactg ctgccaggct ccccaacacc tgaagagccc atttaaggat ccctcgagag
     4021 atcagtgtag cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg
     4081 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta
     4141 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     4201 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     4261 cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc
     4321 ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc
     4381 ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc
     4441 gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg
     4501 cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag
     4561 atccaagctg tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg
     4621 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4681 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
     4741 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4801 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4861 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4921 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4981 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5041 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5101 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5161 ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt
     5221 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5281 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5341 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5401 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     5461 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     5521 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     5581 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     5641 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     5701 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     5761 tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct
     5821 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     5881 cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     5941 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6001 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6061 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6121 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     6181 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6241 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     6301 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6361 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6421 gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat
     6481 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6541 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6601 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6661 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6721 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6781 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6841 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6901 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6961 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7021 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7081 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7141 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7201 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7261 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7321 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7381 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7441 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7501 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7561 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7621 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7681 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7741 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7801 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7861 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7921 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7981 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8041 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8101 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8161 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8221 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8281 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8341 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8401 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8461 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8521 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8581 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8641 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     8701 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     8761 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     8821 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     8881 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     8941 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     9001 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     9061 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
