LOCUS       V023823                 8961 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023823
VERSION     V023823
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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             1660..4425
                     /label="Stat2(NM_019963)"
                     /note="Stat2(NM_019963)"
                     /gene="Stat2"
     polyA_signal    4528..4752
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4798..5226
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5240..5569
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5636..6430
                     /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    6604..6725
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6774..6790)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6798..6814
                     /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(6822..6852)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6867..6888
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7176..7761)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7932..8792)
                     /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(8793..8897)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tggcgcagtg ggagatgttg
     1681 cagaaccttg acagcctgtt cctggaccag ttacaccagg tctactcaca gagcatcctg
     1741 ccaatggacg ttcgacagca tttggctacc tggattgaag accagaactg gagggaagct
     1801 gcgctgggca gtgatgacgc caaggccaac atgctatact tcagcatcct ggaccaactg
     1861 aaccagtggg accactacag ctcagactcc aatcactttt tattacagca taacttgcga
     1921 aaattcagcc gggatattca gacctttccc aatggcccta cccagttggc tgagatgatc
     1981 tttaatcttc ttctggaaga gcaaaggatt ctgaatcagg ctcaaagagc tcaagaggtg
     2041 cagcccccac cagcccccga agcagttgtg gagagccagc agcttgagat tgaaaatcga
     2101 atccagggtt tacatgtgga cattgagttc ttggtgagat ccatcaggca gctgaaggac
     2161 gaacaggatg tcttcagctt cagatacaca gttttcagtc tgaagaagac gtcgtcttca
     2221 gacccccatc agagccaaca ggcgcagctt gtgcaggcaa cagccaacaa agtcgacaga
     2281 atgagaaagg aggtgctgga catctccaaa ggactggttg gccgattaac caccctggtc
     2341 gacctattgc tgcccaagct ggacgagtgg aaggtgcagc agcagaagtc ctgcattgga
     2401 gccccgcccc ccgagctgca gctggaacag ctggaacagt ggttgacggc tggagcaaag
     2461 ttcttgttcc accttcggca gctactgaag cagctgaagg agatgagtca catgcttcgg
     2521 tataagggtg acatgtttgg ccaaggggtg gacctgcaga atgcccaagt catggagtta
     2581 cttcagcgtc tgctccaaag gtcctttgta gtagaaaccc agccctgcat gccccagact
     2641 ctccaccgac ccctcatcct gaagactggg aacaagttca ccgtccgaac aagacttctg
     2701 gtgagactcc aggaaggcag cgaatcactc aaagcagaag tctctgtcga caggaattca
     2761 gacttaccag gcttccggaa gttcaacatt ctgacctcaa accaaaaaac cttgaccccg
     2821 gaggagggcc agagacaggg cttaatttgg gacttcggct tcttgactct ggtggagcaa
     2881 cgcgctgtgg gtgcaggaaa gggcaacaag gggccgctgg cagtgacaga ggagttacat
     2941 gtcatcagct tcgtggttga gtacgtgtac cagggtctga agatgaagct gcagacggac
     3001 actctccctg ttgtgattat ttctaacatg aaccaactct ccattgcctg ggcctccatt
     3061 ctctggttca acatgctcag cccaaatccc aagaaccagc agttcttctg ccaagctccg
     3121 aaagccccct ggagtttgct gggacctgtg ctcagctggc agttctcctc ttacgttggc
     3181 cgaggcctcg attctgagca gctgggcatg ctgagaacca agctgtttgg aaagagctgc
     3241 aagatggagg atgcgctgtt gtcctgggta gacttttgca agcgagagag cccccctggt
     3301 aagatccctt tctggacctg gctggacaaa attctggagc tggtgcacga ccacctgaag
     3361 gatctctgga aggatgggcg catcatgggc tttgtgagcc gcaaccagga acgcagactg
     3421 ctgaagaaga tgttgtctgg caccttccta ctgcgcttca gtgaaacttc tgaagggggc
     3481 attacttgtt cttgggtgga gcaccaggat gacgataaag tcgaaatcta ctcagtgcag
     3541 ccctacacca aggaagtgtt acagtcactc ccactgacag agatcatccg ccactaccag
     3601 gttcttgccg aagagaacat ccccgagaac ccactccgct tcctctatcc ccgaatccct
     3661 cgggacgaag cttttgggtg ttactaccag gaaaaagtta attttgaaga acagaggaaa
     3721 tatttgaaac ataaactcat tgtgatctct aacagacagg tggacgagct gcagcagcct
     3781 ctggagctca aacaggattc agagtcctta gaagtgaatg cagagctctt gttagcacac
     3841 gaccaggagt tgccattgat gatgcagact gggctggttc tgggcacaga gctgaaagtg
     3901 gaccccatac tgagtacagc cccacaagtc ctgctggagc cagccccaca agtcctgctg
     3961 gagccagccc cacaagtccc gctggagcca gccccacaag tcctgctgga gccagcccca
     4021 caagtcctgc tggagccagc cccacaagtc ctgctggagc cagccccaca agtcctgctg
     4081 gagccagccc cacaagtcca gctggagcca gccccacaag tcctgctgga gctagcccca
     4141 caagtcctgc tggagccagc cccacaagtc ctgctggagc tagccccaca agtccagctg
     4201 gagccagcac acttgctgca gcagccatca gagtcagatt tgcctgagga cctgcagcag
     4261 attagcgtgg aggatctgaa aaaattaagc aacccgagta ctgaatacat cacaaccaac
     4321 gaaaacccaa tgttggctgg cgagagcagt ggagatgaga cttccatacc ctaccacagc
     4381 cattttgatg cggatggact cttgggatgg actcttgata ccttctgagg atccactagt
     4441 ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc
     4501 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4561 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4621 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4681 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4741 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     4801 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4861 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4921 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4981 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5041 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5101 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5161 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5221 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5281 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5341 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5401 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5461 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     5521 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     5581 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     5641 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5701 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5761 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     5821 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5881 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5941 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     6001 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     6061 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     6121 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     6181 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6241 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6301 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6361 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6421 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6481 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6541 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6601 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6721 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     6781 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6841 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6901 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6961 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7681 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     7741 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7801 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7861 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7921 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8221 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8521 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8761 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8941 gatctcccga tcccctatgg t
//
