LOCUS       V032738                 9027 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032738
VERSION     V032738
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..3952
                     /label="ADGRG5(NM_153837)"
                     /note="ADGRG5(NM_153837)"
                     /gene="ADGRG5"
     promoter        3997..4208
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4221..4489
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4516..5115
                     /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    5122..5710
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5784..6017
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6089..6210
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6229..6364
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6402..6418)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6426..6442
                     /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(6450..6480)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6495..6516
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6804..7392)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7563..8423)
                     /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(8424..8528)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9002..9018
                     /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 ggatcactgt ggtgcccttt tcctgtgcct
     2401 gtgccttctg actttgcaga atgcaacaac agagacatgg gaagaactcc tgagctacat
     2461 ggagaatatg caggtgtcca ggggccggag ctcagttttt tcctctcgtc aactccacca
     2521 gctggagcag atgctactga acaccagctt cccaggctac aacctgacct tgcagacacc
     2581 caccatccag tctctggcct tcaagctgag ctgtgacttc tctggcctct cgctgaccag
     2641 tgccactctg aagcgggtgc cccaggcagg aggtcagcat gcccggggtc agcacgccat
     2701 gcagttcccc gccgagctga cccgggacgc ctgcaagacc cgccccaggg agctgcggct
     2761 catctgtatc tacttctcca acacccactt tttcaaggat gaaaacaact catctctgct
     2821 gaataactac gtcctggggg cccagctgag tcatgggcac gtgaacaacc tcagggatcc
     2881 tgtgaacatc agcttctggc acaaccaaag cctggaaggc tacaccctga cctgtgtctt
     2941 ctggaaggag ggagccagga aacagccctg ggggggctgg agccctgagg gctgtcgtac
     3001 agagcagccc tcccactctc aggtgctctg ccgctgcaac cacctcacct actttgctgt
     3061 tctcatgcaa ctctccccag ccctggtccc tgcagagttg ctggcacctc ttacgtacat
     3121 ctccctcgtg ggctgcagca tctccatcgt ggcctcgctg atcacagtcc tgctgcactt
     3181 ccatttcagg aagcagagtg actccttaac acgcatccac atgaacctgc atgcctccgt
     3241 gctgctcctg aacatcgcct tcctgctgag ccccgcattc gcaatgtctc ctgtgcccgg
     3301 gtcagcatgc acggctctgg ccgctgccct gcactacgcg ctgctcagct gcctcacctg
     3361 gatggccatc gagggcttca acctctacct cctcctcggg cgtgtctaca acatctacat
     3421 ccgcagatat gtgttcaagc ttggtgtgct aggctggggg gccccagccc tcctggtgct
     3481 gctttccctc tctgtcaaga gctcggtata cggaccctgc acaatccccg tcttcgacag
     3541 ctgggagaat ggcacaggct tccagaacat gtccatatgc tgggtgcgga gccccgtggt
     3601 gcacagtgtc ctggtcatgg gctacggcgg cctcacgtcc ctcttcaacc tggtggtgct
     3661 ggcctgggcg ctgtggaccc tgcgcaggct gcgggagcgg gcggatgcac caagtgtcag
     3721 ggcctgccat gacactgtca ctgtgctggg cctcaccgtg ctgctgggaa ccacctgggc
     3781 cttggccttc ttttcttttg gcgtcttcct gctgccccag ctgttcctct tcaccatctt
     3841 aaactcgctc tacggtttct tccttttcct gtggttctgc tcccagcggt gccgctcaga
     3901 agcagaggcc aaggcacaga tagaggcctt cagctcctcc caaacaacac agtaggcggc
     3961 cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     4021 tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg
     4081 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     4141 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     4201 gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct
     4261 gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct
     4321 gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg
     4381 cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg
     4441 ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac
     4501 cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4561 gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4621 accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4681 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4741 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4801 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     4861 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4921 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     4981 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5041 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5101 aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     5161 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5221 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5281 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5341 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5401 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5461 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5521 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5581 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5641 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5701 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     5761 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct
     5821 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5881 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5941 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6001 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6061 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     6121 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6181 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     6241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6301 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6361 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg
     6421 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6481 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6541 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7201 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8521 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8581 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     8641 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     8701 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     8761 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     8821 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     8881 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     8941 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     9001 tgtaaaacga cggccagtgc caagctg
//
