LOCUS       V039225                 7098 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039225
VERSION     V039225
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..1901
                     /label="IRF1(NM_002198)"
                     /note="IRF1(NM_002198)"
                     /gene="IRF1"
     CDS             1941..2660
                     /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    2698..2922
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2968..3396
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3410..3739
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3806..4600
                     /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    4774..4895
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4944..4960)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4968..4984
                     /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(4992..5022)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5037..5058
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5346..5931)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6102..6962)
                     /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(6963..7067)
                     /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 gccaccatgc ccatcactcg gatgcgcatg agaccctggc
      961 tagagatgca gattaattcc aaccaaatcc cggggctcat ctggattaat aaagaggaga
     1021 tgatcttcca gatcccatgg aagcatgctg ccaagcatgg ctgggacatc aacaaggatg
     1081 cctgtttgtt ccggagctgg gccattcaca caggccgata caaagcaggg gaaaaggagc
     1141 cagatcccaa gacgtggaag gccaactttc gctgtgccat gaactccctg ccagatatcg
     1201 aggaggtgaa agaccagagc aggaacaagg gcagctcagc tgtgcgagtg taccggatgc
     1261 ttccacctct caccaagaac cagagaaaag aaagaaagtc gaagtccagc cgagatgcta
     1321 agagcaaggc caagaggaag tcatgtgggg attccagccc tgataccttc tctgatggac
     1381 tcagcagctc cactctgcct gatgaccaca gcagctacac agttccaggc tacatgcagg
     1441 acttggaggt ggagcaggcc ctgactccag cactgtcgcc atgtgctgtc agcagcactc
     1501 tccccgactg gcacatccca gtggaagttg tgccggacag caccagtgat ctgtacaact
     1561 tccaggtgtc acccatgccc tccacctctg aagctacaac agatgaggat gaggaaggga
     1621 aattacctga ggacatcatg aagctcttgg agcagtcgga gtggcagcca acaaacgtgg
     1681 atgggaaggg gtacctactc aatgaacctg gagtccagcc cacctctgtc tatggagact
     1741 ttagctgtaa ggaggagcca gaaattgaca gcccaggggg ggatattggg ctgagtctac
     1801 agcgtgtctt cacagatctg aagaacatgg atgccacctg gctggacagc ctgctgaccc
     1861 cagtccggtt gccctccatc caggccattc cctgtgcacc gggctcgagt ggcggtggag
     1921 gaagcggcgg aggtggatct atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1981 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2041 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2101 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2161 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2221 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2281 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2341 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2401 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2461 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2521 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2581 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2641 tggacgagct gtacaagtag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     2701 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2761 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2821 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2881 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2941 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3001 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3061 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3121 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3181 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3241 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3301 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3361 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3421 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3481 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3541 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3601 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3661 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3721 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3781 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3841 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3901 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3961 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4021 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4081 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4141 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4201 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4261 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4321 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4381 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4441 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4501 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4561 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4621 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4681 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4741 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4801 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4861 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4921 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4981 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5041 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5101 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5161 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5221 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5281 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5341 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5401 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5461 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5521 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5581 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5641 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5701 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5761 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5821 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5881 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5941 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6001 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6061 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6181 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6541 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6961 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7021 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7081 aaagtgccac ctgacgtc
//
