LOCUS       V028217                 8471 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028217
VERSION     V028217
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..3396
                     /label="GPR68(NM_001177676)"
                     /note="GPR68(NM_001177676)"
                     /gene="GPR68"
     promoter        3441..3652
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3665..3933
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3960..4559
                     /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    4566..5154
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5228..5461
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5533..5654
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5673..5808
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5846..5862)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5870..5886
                     /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(5894..5924)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5939..5960
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6248..6836)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7007..7867)
                     /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(7868..7972)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8446..8462
                     /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 catggggaac atcactgcag acaactcctc gatgagctgt
     2341 accatcgacc ataccatcca ccagacgctg gccccggtgg tctatgttac cgtgctggtg
     2401 gtgggcttcc cggccaactg cctgtccctc tacttcggct acctgcagat caaggcccgg
     2461 aacgagctgg gcgtgtacct gtgcaacctg acggtggccg acctcttcta catctgctcg
     2521 ctgcccttct ggctgcagta cgtgctgcag cacgacaact ggtctcacgg cgacctgtcc
     2581 tgccaggtgt gcggcatcct cctgtacgag aacatctaca tcagcgtggg cttcctctgc
     2641 tgcatctccg tggaccgcta cctggctgtg gcccatccct tccgcttcca ccagttccgg
     2701 accctgaagg cggccgtcgg cgtcagcgtg gtcatctggg ccaaggagct gctgaccagc
     2761 atctacttcc tgatgcacga ggaggtcatc gaggacgaga accagcaccg cgtgtgcttt
     2821 gagcactacc ccatccaggc atggcagcgc gccatcaact actaccgctt cctggtgggc
     2881 ttcctcttcc ccatctgcct gctgctggcg tcctaccagg gcatcctgcg cgccgtgcgc
     2941 cggagccacg gcacccagaa gagccgcaag gaccagatcc agcggctggt gctcagcacc
     3001 gtggtcatct tcctggcctg cttcctgccc taccacgtgt tgctgctggt gcgcagcgtc
     3061 tgggaggcca gctgcgactt cgccaagggc gttttcaacg cctaccactt ctccctcctg
     3121 ctcaccagct tcaactgcgt cgccgacccc gtgctctact gcttcgtcag cgagaccacc
     3181 caccgggacc tggcccgcct ccgcggggcc tgcctggcct tcctcacctg ctccaggacc
     3241 ggccgggcca gggaggccta cccgctgggt gcccccgagg cctccgggaa aagcggggcc
     3301 cagggtgagg agcccgagct gttgaccaag ctccacccgg ccttccagac ccctaactcg
     3361 ccagggtcgg gcgggttccc cacgggcagg ttggcctagg cggccgcgaa ggatctgcga
     3421 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     3481 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     3541 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     3601 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     3661 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     3721 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     3781 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     3841 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     3901 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga
     3961 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     4021 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     4081 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4141 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     4201 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     4261 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     4321 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     4381 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     4441 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     4501 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     4561 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4621 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4681 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4741 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4801 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4861 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4921 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4981 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5041 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5101 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta
     5161 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     5221 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     5281 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5341 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5401 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5461 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     5521 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5581 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5641 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     5701 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5761 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     5821 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5881 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5941 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6001 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6061 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6121 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6181 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6241 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6301 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6361 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6421 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6481 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6541 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6601 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6661 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6721 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6781 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6961 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7021 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7081 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7141 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7201 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7261 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7321 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7381 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7441 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7501 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7561 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7621 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7681 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7741 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7801 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7861 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7921 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7981 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8041 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8101 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8161 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     8221 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     8281 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     8341 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     8401 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     8461 gtgccaagct g
//
