LOCUS       V037269                 9247 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037269
VERSION     V037269
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             2303..3343
                     /label="JUNB(NM_002229)"
                     /note="JUNB(NM_002229)"
                     /gene="JUNB"
     CDS             3344..3361
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        3406..3617
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3630..3898
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3927..3936
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3957..4619
                     /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             4689..4742
                     /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             4743..5342
                     /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    5343..5930
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6004..6237
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6309..6430
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6449..6584
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6622..6638)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6646..6662
                     /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(6670..6700)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6715..6736
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7024..7612)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7783..8643)
                     /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(8644..8748)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9222..9238
                     /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 gaattcgcca ccatgtgcac taaaatggaa cagcccttct accacgacga
     2341 ctcatacaca gctacgggat acggccgggc ccctggtggc ctctctctac acgactacaa
     2401 actcctgaaa ccgagcctgg cggtcaacct ggccgacccc taccggagtc tcaaagcgcc
     2461 tggggctcgc ggacccggcc cagagggcgg cggtggcggc agctactttt ctggtcaggg
     2521 ctcggacacc ggcgcgtctc tcaagctcgc ctcttcggag ctggaacgcc tgattgtccc
     2581 caacagcaac ggcgtgatca cgacgacgcc tacacccccg ggacagtact tttacccccg
     2641 cgggggtggc agcggtggag gtgcaggggg cgcagggggc ggcgtcaccg aggagcagga
     2701 gggcttcgcc gacggctttg tcaaagccct ggacgatctg cacaagatga accacgtgac
     2761 accccccaac gtgtccctgg gcgctaccgg ggggcccccg gctgggcccg ggggcgtcta
     2821 cgccggcccg gagccacctc ccgtttacac caacctcagc agctactccc cagcctctgc
     2881 gtcctcggga ggcgccgggg ctgccgtcgg gaccgggagc tcgtacccga cgaccaccat
     2941 cagctacctc ccacacgcgc cgcccttcgc cggtggccac ccggcgcagc tgggcttggg
     3001 ccgcggcgcc tccaccttca aggaggaacc gcagaccgtg ccggaggcgc gcagccggga
     3061 cgccacgccg ccggtgtccc ccatcaacat ggaagaccaa gagcgcatca aagtggagcg
     3121 caagcggctg cggaaccggc tggcggccac caagtgccgg aagcggaagc tggagcgcat
     3181 cgcgcgcctg gaggacaagg tgaagacgct caaggccgag aacgcggggc tgtcgagtac
     3241 cgccggcctc ctccgggagc aggtggccca gctcaaacag aaggtcatga cccacgtcag
     3301 caacggctgt cagctgctgc ttggggtcaa gggacacgcc ttccatcatc accatcacca
     3361 ttgagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     3421 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     3481 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     3541 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     3601 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     3661 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     3721 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     3781 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     3841 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     3901 agctgtgacc ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg
     3961 ccatggagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg
     4021 gcggcggaga gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag
     4081 gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact
     4141 tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct
     4201 acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct
     4261 accgctacga ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg
     4321 aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc
     4381 accccatggg cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg
     4441 gcggctacta cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca
     4501 gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca
     4561 acaccgagct gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca
     4621 gatcccgcgc tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg
     4681 gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     4741 ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     4801 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     4861 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4921 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4981 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5041 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     5101 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5161 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     5221 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5281 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     5341 gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5401 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5461 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5521 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5581 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5641 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5701 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     5761 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5821 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5881 tccgccttcg ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt
     5941 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6001 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
     6061 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6121 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6181 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6241 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6301 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6361 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6421 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
     6481 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6541 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     6601 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6661 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6721 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6781 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6841 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6901 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6961 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7441 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7741 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7801 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7861 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7921 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7981 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8041 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8101 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8161 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8221 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8281 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8341 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8401 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8461 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8521 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8581 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8641 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8701 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8761 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8821 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8881 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     8941 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9001 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     9061 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     9121 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     9181 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9241 caagctg
//
