LOCUS       V023027                 9124 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023027
VERSION     V023027
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..1422
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1471..3780
                     /label="STAT3(NM_139276)"
                     /note="STAT3(NM_139276)"
                     /gene="STAT3"
     CDS             3787..3876
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3900..4488
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4554..4602
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4804..5232
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5246..5575
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5642..6436
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6612..6733
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6782..6798)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6806..6822
                     /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(6830..6860)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6875..6896
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7184..7772)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7943..8803)
                     /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(8804..8908)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggt gagcaagggc
      721 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      781 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      841 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      901 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      961 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1021 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1081 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1141 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1201 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1261 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1321 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1381 accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga
     1441 ggcggcggag ggagtggagg agggggaagc atggcccaat ggaatcagct acagcagctt
     1501 gacacacggt acctggagca gctccatcag ctctacagtg acagcttccc aatggagctg
     1561 cggcagtttc tggccccttg gattgagagt caagattggg catatgcggc cagcaaagaa
     1621 tcacatgcca ctttggtgtt tcataatctc ctgggagaga ttgaccagca gtatagccgc
     1681 ttcctgcaag agtcgaatgt tctctatcag cacaatctac gaagaatcaa gcagtttctt
     1741 cagagcaggt atcttgagaa gccaatggag attgcccgga ttgtggcccg gtgcctgtgg
     1801 gaagaatcac gccttctaca gactgcagcc actgcggccc agcaaggggg ccaggccaac
     1861 caccccacag cagccgtggt gacggagaag cagcagatgc tggagcagca ccttcaggat
     1921 gtccggaaga gagtgcagga tctagaacag aaaatgaaag tggtagagaa tctccaggat
     1981 gactttgatt tcaactataa aaccctcaag agtcaaggag acatgcaaga tctgaatgga
     2041 aacaaccagt cagtgaccag gcagaagatg cagcagctgg aacagatgct cactgcgctg
     2101 gaccagatgc ggagaagcat cgtgagtgag ctggcggggc ttttgtcagc gatggagtac
     2161 gtgcagaaaa ctctcacgga cgaggagctg gctgactgga agaggcggca acagattgcc
     2221 tgcattggag gcccgcccaa catctgccta gatcggctag aaaactggat aacgtcatta
     2281 gcagaatctc aacttcagac ccgtcaacaa attaagaaac tggaggagtt gcagcaaaaa
     2341 gtttcctaca aaggggaccc cattgtacag caccggccga tgctggagga gagaatcgtg
     2401 gagctgttta gaaacttaat gaaaagtgcc tttgtggtgg agcggcagcc ctgcatgccc
     2461 atgcatcctg accggcccct cgtcatcaag accggcgtcc agttcactac taaagtcagg
     2521 ttgctggtca aattccctga gttgaattat cagcttaaaa ttaaagtgtg cattgacaaa
     2581 gactctgggg acgttgcagc tctcagagga tcccggaaat ttaacattct gggcacaaac
     2641 acaaaagtga tgaacatgga agaatccaac aacggcagcc tctctgcaga attcaaacac
     2701 ttgaccctga gggagcagag atgtgggaat gggggccgag ccaattgtga tgcttccctg
     2761 attgtgactg aggagctgca cctgatcacc tttgagaccg aggtgtatca ccaaggcctc
     2821 aagattgacc tagagaccca ctccttgcca gttgtggtga tctccaacat ctgtcagatg
     2881 ccaaatgcct gggcgtccat cctgtggtac aacatgctga ccaacaatcc caagaatgta
     2941 aactttttta ccaagccccc aattggaacc tgggatcaag tggccgaggt cctgagctgg
     3001 cagttctcct ccaccaccaa gcgaggactg agcatcgagc agctgactac actggcagag
     3061 aaactcttgg gacctggtgt gaattattca gggtgtcaga tcacatgggc taaattttgc
     3121 aaagaaaaca tggctggcaa gggcttctcc ttctgggtct ggctggacaa tatcattgac
     3181 cttgtgaaaa agtacatcct ggccctttgg aacgaagggt acatcatggg ctttatcagt
     3241 aaggagcggg agcgggccat cttgagcact aagcctccag gcaccttcct gctaagattc
     3301 agtgaaagca gcaaagaagg aggcgtcact ttcacttggg tggagaagga catcagcggt
     3361 aagacccaga tccagtccgt ggaaccatac acaaagcagc agctgaacaa catgtcattt
     3421 gctgaaatca tcatgggcta taagatcatg gatgctacca atatcctggt gtctccactg
     3481 gtctatctct atcctgacat tcccaaggag gaggcattcg gaaagtattg tcggccagag
     3541 agccaggagc atcctgaagc tgacccaggt agcgctgccc catacctgaa gaccaagttt
     3601 atctgtgtga caccaacgac ctgcagcaat accattgacc tgccgatgtc cccccgcact
     3661 ttagattcat tgatgcagtt tggaaataat ggtgaaggtg ctgaaccctc agcaggaggg
     3721 cagtttgagt ccctcacctt tgacatggag ttgacctcgg agtgcgctac ctcccccatg
     3781 ctcgaggaac aaaaactaat atcagaggaa gatttggaac agaaactaat cagtgaagaa
     3841 gatttagagc aaaagttaat ttccgaggag gacttatagt gagggcccga tatctcgaca
     3901 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3961 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4021 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4081 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4141 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4201 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4261 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     4321 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4381 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4441 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg
     4501 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4561 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4621 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4681 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4741 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     4801 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat
     4981 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5101 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5161 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5221 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5281 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5401 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5461 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     5521 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5581 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     5641 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5701 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5761 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5821 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5881 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5941 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     6001 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     6061 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     6121 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     6181 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     6241 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     6301 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6361 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6421 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     6481 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6541 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6601 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6721 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6781 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6841 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6901 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6961 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7021 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7081 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7141 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7201 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7261 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7321 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     7381 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7441 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7501 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7561 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7621 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7681 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7741 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7801 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7861 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7921 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7981 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8041 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8101 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8161 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8221 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8281 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8341 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8401 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8461 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8521 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8581 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8641 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8701 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8761 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8881 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8941 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
     9001 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     9061 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     9121 tagg
//
