LOCUS       V023065                 9756 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023065
VERSION     V023065
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             2301..4610
                     /label="Stat3(NM_213659)"
                     /note="Stat3(NM_213659)"
                     /gene="Stat3"
     CDS             4617..4682
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4726..4937
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4950..5218
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5245..5844
                     /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    5851..6439
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6513..6746
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6818..6939
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6958..7093
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7131..7147)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7155..7171
                     /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(7179..7209)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7224..7245
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7533..8121)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8292..9152)
                     /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(9153..9257)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9731..9747
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atggctcagt ggaaccagct gcagcagctg gacacacgct
     2341 acctggagca gctgcaccag ctgtacagcg acagcttccc catggagctg cggcagttcc
     2401 tggcaccttg gattgagagt caagactggg catatgcagc cagcaaagag tcacatgcca
     2461 cgttggtgtt tcataatctc ttgggtgaaa ttgaccagca atatagccga ttcctgcaag
     2521 agtccaatgt cctctatcag cacaaccttc gaagaatcaa gcagtttctg cagagcaggt
     2581 atcttgagaa gccaatggaa attgcccgga tcgtggcccg atgcctgtgg gaagagtctc
     2641 gcctcctcca gacggcagcc acggcagccc agcaaggggg ccaggccaac cacccaacag
     2701 ccgccgtagt gacagagaag cagcagatgt tggagcagca tcttcaggat gtccggaagc
     2761 gagtgcagga tctagaacag aaaatgaagg tggtggagaa cctccaggac gactttgatt
     2821 tcaactacaa aaccctcaag agccaaggag acatgcagga tctgaatgga aacaaccagt
     2881 ctgtgaccag acagaagatg cagcagctgg aacagatgct cacagccctg gaccagatgc
     2941 ggagaagcat tgtgagtgag ctggcggggc tcttgtcagc aatggagtac gtgcagaaga
     3001 cactgactga tgaagagctg gctgactgga agaggcggca gcagatcgcg tgcatcggag
     3061 gccctcccaa catctgcctg gaccgtctgg aaaactggat aacttcatta gcagaatctc
     3121 aacttcagac ccgccaacaa attaagaaac tggaggagct gcagcagaaa gtgtcctaca
     3181 agggcgaccc tatcgtgcag caccggccca tgctggagga gaggatcgtg gagctgttca
     3241 gaaacttaat gaagagtgcc ttcgtggtgg agcggcagcc ctgcatgccc atgcacccgg
     3301 accggccctt agtcatcaag actggtgtcc agtttaccac gaaagtcagg ttgctggtca
     3361 aatttcctga gttgaattat cagcttaaaa ttaaagtgtg cattgataaa gactctgggg
     3421 atgttgctgc cctcagaggg tctcggaaat ttaacattct gggcacgaac acaaaagtga
     3481 tgaacatgga ggagtctaac aacggcagcc tgtctgcaga gttcaagcac ctgaccctta
     3541 gggagcagag atgtgggaat ggaggccgtg ccaattgtga tgcctccttg atcgtgactg
     3601 aggagctgca cctgatcacc ttcgagactg aggtgtacca ccaaggcctc aagattgacc
     3661 tagagaccca ctccttgcca gttgtggtga tctccaacat ctgtcagatg ccaaatgctt
     3721 gggcatcaat cctgtggtat aacatgctga ccaataaccc caagaacgtg aacttcttca
     3781 ctaagccgcc aattggaacc tgggaccaag tggccgaggt gctcagctgg cagttctcgt
     3841 ccaccaccaa gcgggggctg agcatcgagc agctgacaac gctggctgag aagctcctag
     3901 ggcctggtgt gaactactca gggtgtcaga tcacatgggc taaattctgc aaagaaaaca
     3961 tggctggcaa gggcttctcc ttctgggtct ggctagacaa tatcatcgac cttgtgaaaa
     4021 agtatatctt ggccctttgg aatgaagggt acatcatggg tttcatcagc aaggagcggg
     4081 agcgggccat cctaagcaca aagcccccgg gcaccttcct actgcgcttc agcgagagca
     4141 gcaaagaagg aggggtcact ttcacttggg tggaaaagga catcagtggc aagacccaga
     4201 tccagtctgt agagccatac accaagcagc agctgaacaa catgtcattt gctgaaatca
     4261 tcatgggcta taagatcatg gatgcgacca acatcctggt gtctccactt gtctacctct
     4321 accccgacat tcccaaggag gaggcatttg gaaagtactg taggcccgag agccaggagc
     4381 accccgaagc cgacccaggt agtgctgccc cgtacctgaa gaccaagttc atctgtgtga
     4441 caccaacgac ctgcagcaat accattgacc tgccgatgtc cccccgcact ttagattcat
     4501 tgatgcagtt tggaaataac ggtgaaggtg ctgagccctc agcaggaggg cagtttgagt
     4561 cgctcacgtt tgacatggat ctgacctcgg agtgtgctac ctcccccatg ggatccgact
     4621 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     4681 agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     4741 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     4801 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4861 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4921 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     4981 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     5041 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     5101 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     5161 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     5221 agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc
     5281 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     5341 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     5401 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     5461 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     5521 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     5581 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     5641 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     5701 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5761 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5821 atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa
     5881 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5941 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     6001 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6061 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6121 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6181 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6241 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6301 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6361 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6421 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     6481 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa
     6541 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6601 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6661 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6721 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6781 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6841 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6901 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6961 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     7021 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     7081 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg
     7141 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8221 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8281 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8341 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8401 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8461 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8521 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8581 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8641 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8701 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8761 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8821 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8881 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8941 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9001 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9061 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9121 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9181 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9241 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     9301 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     9361 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     9421 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     9481 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     9541 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     9601 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9661 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9721 tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
