LOCUS       V030402                 9007 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030402
VERSION     V030402
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..3861
                     /label="USP3(NM_006537)"
                     /note="USP3(NM_006537)"
                     /gene="USP3"
     CDS             3868..3933
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3977..4188
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4201..4469
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4496..5095
                     /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    5102..5690
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5764..5997
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6069..6190
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6209..6344
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6382..6398)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6406..6422
                     /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(6430..6460)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6475..6496
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6784..7372)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7543..8403)
                     /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(8404..8508)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8982..8998
                     /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 catggagtgt ccacacctga gctccagcgt ctgcattgct
     2341 ccggactcag ccaagttccc caacggctcc ccgtcgtcct ggtgctgcag cgtgtgccgg
     2401 tccaacaaaa gcccttgggt ctgtttgact tgttcaagtg tccactgtgg aaggtatgtg
     2461 aatggccatg caaaaaaaca ttatgaagat gcacaagtac ctttaaccaa ccataagaaa
     2521 tcagaaaagc aagataaagt tcagcacaca gtatgtatgg attgcagtag ctacagtaca
     2581 tactgttatc gctgtgatga ttttgtggtt aatgacacca agctgggact ggtacagaaa
     2641 gtcagagaac acttacagaa cttggaaaac tcagctttca cagctgacag gcataagaaa
     2701 agaaaacttt tggaaaactc aacactaaac agcaagttat taaaagtaaa tggaagcacc
     2761 actgccattt gtgccacagg ccttcggaat ttggggaaca catgtttcat gaatgccatc
     2821 cttcagtcac tcagtaacat tgagcagttt tgctgttatt tcaaagaact gcccgccgtg
     2881 gagttaagga atgggaaaac agcaggaagg cggacatacc acaccaggag ccaaggggat
     2941 aacaatgtgt ctttggtaga agagtttaga aagacactct gtgctttatg gcaaggcagc
     3001 cagactgcat ttagcccaga gtccttattt tatgttgttt ggaagattat gccaaacttt
     3061 aggggctatc aacagcagga cgcccatgaa ttcatgcgct accttttgga ccacctacac
     3121 ttggaacttc agggcggttt caacggtgtt tcccgctcag caattctgca ggagaattct
     3181 actctgtctg caagtaacaa gtgttgcata aatggagcat ctactgttgt cacggctata
     3241 ttcggaggca ttctccaaaa tgaggttaac tgcctcatat gtgggacaga atctagaaag
     3301 tttgatccat tcctagacct ttcattagat attccaagtc agttcagaag taagcgctct
     3361 aagaatcaag aaaatggacc agtttgttcg ttacgagatt gtcttcgcag ttttaccgac
     3421 ttagaagaac ttgatgagac agagttatat atgtgccata aatgcaaaaa gaaacaaaag
     3481 tccacaaaaa agttttggat tcaaaaacta cccaaggtgc tatgcttaca tttgaaaaga
     3541 tttcattgga cagcatattt aagaaataaa gttgatacat acgtagaatt tccactgaga
     3601 ggcctagaca tgaaatgcta cttactagag cctgagaaca gtggcccgga gagctgcctg
     3661 tatgacctcg ccgctgtggt ggtgcaccat ggttccgggg ttggttctgg acattacaca
     3721 gcatacgcaa ctcacgaagg ccgctggttc cacttcaatg acagtactgt aacactgact
     3781 gacgaggaga ctgtggtgaa ggcgaaggcc tacatccttt tctacgtgga acaccaggcc
     3841 aaagctggat cggataaact tggatccgac tacaaagacc atgacggtga ttataaagat
     3901 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc
     3961 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     4021 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     4081 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     4141 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     4201 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     4261 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     4321 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     4381 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     4441 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac
     4501 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac
     4561 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca
     4621 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4681 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4741 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4801 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4861 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4921 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4981 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5041 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga
     5101 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5161 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5221 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5281 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5341 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5401 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5461 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5521 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5581 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5641 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt
     5701 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5761 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
     5821 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5881 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5941 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6001 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6061 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6181 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
     6241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     6361 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6421 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6481 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6541 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7201 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8521 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8581 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8641 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     8701 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     8761 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     8821 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     8881 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     8941 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9001 caagctg
//
