LOCUS       V042653                 8676 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042653
VERSION     V042653
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             927..3479
                     /label="STAT2(NM_005419)"
                     /note="STAT2(NM_005419)"
                     /gene="STAT2"
     CDS             3519..4238
                     /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"
     polyA_signal    4276..4500
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4546..4974
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4988..5317
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5384..6178
                     /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    6352..6473
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6522..6538)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6546..6562
                     /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(6570..6600)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6615..6636
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6924..7509)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7680..8540)
                     /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(8541..8645)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaagcc actgggatcc gccaccatgg cgcagtggga aatgctgcag aatcttgaca
      961 gcccctttca ggatcagctg caccagcttt actcgcacag cctcctgcct gtggacattc
     1021 gacagtactt ggctgtctgg attgaagacc agaactggca ggaagctgca cttgggagtg
     1081 atgattccaa ggctaccatg ctattcttcc acttcttgga tcagctgaac tatgagtgtg
     1141 gccgttgcag ccaggaccca gagtccttgt tgctgcagca caatttgcgg aaattctgcc
     1201 gggacattca gcccttttcc caggatccta cccagttggc tgagatgatc tttaacctcc
     1261 ttctggaaga aaaaagaatt ttgatccagg ctcagagggc ccaattggaa caaggagagc
     1321 cagttctcga aacacctgtg gagagccagc aacatgagat tgaatcccgg atcctggatt
     1381 taagggctat gatggagaag ctggtaaaat ccatcagcca actgaaagac cagcaggatg
     1441 tcttctgctt ccgatataag atccaggcca aagggaagac accctctctg gacccccatc
     1501 agaccaaaga gcagaagatt ctgcaggaaa ctctcaatga actggacaaa aggagaaagg
     1561 aggtgctgga tgcctccaaa gcactgctag gccgattaac taccctaatc gagctactgc
     1621 tgccaaagtt ggaggagtgg aaggcccagc agcaaaaagc ctgcatcaga gctcccattg
     1681 accacgggtt ggaacagctg gagacatggt tcacagctgg agcaaagctg ttgtttcacc
     1741 tgaggcagct gctgaaggag ctgaagggac tgagttgcct ggttagctat caggatgacc
     1801 ctctgaccaa aggggtggac ctacgcaacg cccaggtcac agagttgcta cagcgtctgc
     1861 tccacagagc ctttgtggta gaaacccagc cctgcatgcc ccaaactccc catcgacccc
     1921 tcatcctcaa gactggcagc aagttcaccg tccgaacaag gctgctggtg agactccagg
     1981 aaggcaatga gtcactgact gtggaagtct ccattgacag gaatcctcct caattacaag
     2041 gcttccggaa gttcaacatt ctgacttcaa accagaaaac tttgaccccc gagaaggggc
     2101 agagtcaggg tttgatttgg gactttggtt acctgactct ggtggagcaa cgttcaggtg
     2161 gttcaggaaa gggcagcaat aaggggccac taggtgtgac agaggaactg cacatcatca
     2221 gcttcacggt caaatatacc taccagggtc tgaagcagga gctgaaaacg gacaccctcc
     2281 ctgtggtgat tatttccaac atgaaccagc tctcaattgc ctgggcttca gttctctggt
     2341 tcaatttgct cagcccaaac cttcagaacc agcagttctt ctccaacccc cccaaggccc
     2401 cctggagctt gctgggccct gctctcagtt ggcagttctc ctcctatgtt ggccgaggcc
     2461 tcaactcaga ccagctgagc atgctgagaa acaagctgtt cgggcagaac tgtaggactg
     2521 aggatccatt attgtcctgg gctgacttca ctaagcgaga gagccctcct ggcaagttac
     2581 cattctggac atggctggac aaaattctgg agttggtaca tgaccacctg aaggatctct
     2641 ggaatgatgg acgcatcatg ggctttgtga gtcggagcca ggagcgccgg ctgctgaaga
     2701 agaccatgtc tggcaccttt ctactgcgct tcagtgaatc gtcagaaggg ggcattacct
     2761 gctcctgggt ggagcaccag gatgatgaca aggtgctcat ctactctgtg caaccgtaca
     2821 cgaaggaggt gctgcagtca ctcccgctga ctgaaatcat ccgccattac cagttgctca
     2881 ctgaggagaa tatacctgaa aacccactgc gcttcctcta tccccgaatc ccccgggatg
     2941 aagcttttgg gtgctactac caggagaaag ttaatctcca ggaacggagg aaatacctga
     3001 aacacaggct cattgtggtc tctaatagac aggtggatga actgcaacaa ccgctggagc
     3061 ttaagccaga gccagagctg gagtcattag agctggaact agggctggtg ccagagccag
     3121 agctcagcct ggacttagag ccactgctga aggcagggct ggatctgggg ccagagctag
     3181 agtctgtgct ggagtccact ctggagcctg tgatagagcc cacactatgc atggtatcac
     3241 aaacagtgcc agagccagac caaggacctg tatcacagcc agtgccagag ccagatttgc
     3301 cctgtgatct gagacatttg aacactgagc caatggaaat cttcagaaac tgtgtaaaga
     3361 ttgaagaaat catgccgaat ggtgacccac tgttggctgg ccagaacacc gtggatgagg
     3421 tttacgtctc ccgccccagc cacttctaca ctgatggacc cttgatgcct tctgacttcc
     3481 tctcgagtgg cggtggagga agcggcggag gtggatctat ggtgagcaag ggcgaggagc
     3541 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3601 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3661 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3721 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3781 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3841 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3901 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3961 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     4021 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     4081 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     4141 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     4201 ccgggatcac tctcggcatg gacgagctgt acaagtagtc tagagggccc gtttaaaccc
     4261 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4321 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4381 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4441 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4501 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4561 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4621 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4681 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4741 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4801 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4861 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4921 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4981 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5041 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5101 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5161 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5221 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     5281 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     5341 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5401 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5461 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5521 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5581 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5641 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5701 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5761 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5821 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5881 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5941 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6001 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6061 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6121 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6181 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6241 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6301 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6361 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6421 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6481 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6541 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6601 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6661 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6721 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6781 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6841 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6901 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6961 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7021 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7081 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7141 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7201 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7261 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7321 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7381 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7441 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7501 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7561 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7621 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7681 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7741 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7801 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7861 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7921 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7981 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8041 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8101 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8161 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8221 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8281 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8341 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8401 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8461 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8521 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8581 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8641 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
