LOCUS       V035336                 8947 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035336
VERSION     V035336
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..2973
                     /label="Slx9(NM_133998)"
                     /note="Slx9(NM_133998)"
                     /gene="Slx9"
     CDS             2980..3045
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3089..3300
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3313..3581
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3610..3619
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3640..4302
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4372..4425
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4426..5025
                     /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    5026..5614
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5688..5921
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5993..6114
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6154..6289
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6322..6338)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6346..6362
                     /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(6370..6400)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6415..6436
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6724..7312)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7483..8343)
                     /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(8344..8448)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8922..8938
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg ggaaagtgag ggcgctgcgg
     2341 gctcgtgtgc accgggcggc cgtgagaccc gacggggatt ctgcaccggg ccctgtgccc
     2401 cgcgccgtgg agccggcgct cccacaatcc ccggcgggcg gagcgggggc gaaggactgg
     2461 acattcgttc acaatgacat ctttgccagg acacagatag accctagtgc gttggtgcag
     2521 aggctggagc tggaccagag gagtgtggtg tccctaaaga gaggtgcaga gccaaaggcc
     2581 attttgccta agaaggaaaa gctgaagctt cgccgtgagc gatggttaca gaaaattgaa
     2641 gccataaagc tggcagagca gaagctcagg gaggaacgaa agcgcaaggc catggtggtg
     2701 gtaggcgacc tgcaccccct gagggatgcc ctacctgagc tgcaggaact ggaggccggc
     2761 cgccagcggc agcaagccag acgcagggtg accagcaagc cccggccagt ggagctcagc
     2821 cgaatgacca cagtacagag gcagcagctt cttgaagaag aaaggacccg gtttcagaag
     2881 ctgctggcca gcccaaccta cagagccagt cccctgctgg ccatcgggca gcagctggca
     2941 caccagatgc agctggaagg tggcaaacag ctcggatccg actacaaaga ccatgacggt
     3001 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg
     3061 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     3121 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac
     3181 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     3241 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     3301 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc
     3361 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg
     3421 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct
     3481 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc
     3541 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct
     3601 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc
     3661 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc
     3721 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc
     3781 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc
     3841 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc
     3901 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc
     3961 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc
     4021 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac
     4081 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc
     4141 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg
     4201 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc
     4261 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg
     4321 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga
     4381 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag
     4441 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4501 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4561 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4621 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4681 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4741 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4801 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4861 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4921 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4981 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat
     5041 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5101 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5161 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5221 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5281 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5341 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5401 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5461 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5521 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     5581 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta
     5641 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     5701 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca
     5761 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     5821 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     5881 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     5941 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     6001 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6061 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6121 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6181 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6241 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     6301 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6361 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6421 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6481 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6541 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6721 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7141 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7441 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7501 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7561 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7621 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7681 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7741 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7801 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7861 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7921 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7981 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8041 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8101 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8161 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8221 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8281 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8341 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8401 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8461 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8521 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8581 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     8641 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     8701 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     8761 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     8821 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     8881 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     8941 caagctg
//
