LOCUS       V026111                 7866 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026111
VERSION     V026111
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        236..615
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        616..819
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        864..882
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             965..2668
                     /label="PFKFB3(NM_001363545)"
                     /note="PFKFB3(NM_001363545)"
                     /gene="PFKFB3"
     CDS             2696..3415
                     /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    3467..3691
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3737..4165
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4179..4508
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4575..5369
                     /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    5543..5664
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5713..5729)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5737..5753
                     /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(5761..5791)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5806..5827
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6115..6700)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6871..7731)
                     /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(7732..7836)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc
      961 caccatgccg ttggaactga cgcagagccg agtgcagaag atctgggtgc ccgtggacca
     1021 caggccctcg ttgcccagat cctgtgggcc aaagctgacc aactccccca ccgtcatcgt
     1081 catggtgggc ctccccgccc ggggcaagac ctacatctcc aagaagctga ctcgctacct
     1141 caactggatt ggcgtcccca caaaagtgtt caacgtcggg gagtatcgcc gggaggctgt
     1201 gaagcagtac agctcctaca acttcttccg ccccgacaat gaggaagcca tgaaagtccg
     1261 gaagcaatgt gccttagctg ccttgagaga tgtcaaaagc tacctggcga aagaaggggg
     1321 acaaattgcg gttttcgatg ccaccaatac tactagagag aggagacaca tgatccttca
     1381 ttttgccaaa gaaaatgact ttaaggcgtt tttcatcgag tcggtgtgcg acgaccctac
     1441 agttgtggcc tccaatatca tggaagttaa aatctccagc ccggattaca aagactgcaa
     1501 ctcggcagaa gccatggacg acttcatgaa gaggatcagt tgctatgaag ccagctacca
     1561 gcccctcgac cccgacaaat gcgacaggga cttgtcgctg atcaaggtga ttgacgtggg
     1621 ccggaggttc ctggtgaacc gggtgcagga ccacatccag agccgcatcg tgtactacct
     1681 gatgaacatc cacgtgcagc cgcgtaccat ctacctgtgc cggcacggcg agaacgagca
     1741 caacctccag ggccgcatcg ggggcgactc aggcctgtcc agccggggca agaagtttgc
     1801 cagtgctctg agcaagttcg tggaggagca gaacctgaag gacctgcgcg tgtggaccag
     1861 ccagctgaag agcaccatcc agacggccga ggcgctgcgg ctgccctacg agcagtggaa
     1921 ggcgctcaat gagatcgacg cgggcgtctg tgaggagctg acctacgagg agatcaggga
     1981 cacctaccct gaggagtatg cgctgcggga gcaggacaag tactattacc gctaccccac
     2041 cggggagtcc taccaggacc tggtccagcg cttggagcca gtgatcatgg agctggagcg
     2101 gcaggagaat gtgctggtca tctgccacca ggccgtcctg cgctgcctgc ttgcctactt
     2161 cctggataag agtgcagagg agatgcccta cctgaaatgc cctcttcaca ccgtcctgaa
     2221 actgacgcct gtcgcttatg gctgccgtgt ggaatccatc tacctgaacg tggagtccgt
     2281 ctgcacacac cgggagaggt cagaggatgc aaagaaggga cctaacccgc tcatgagacg
     2341 caatagtgtc accccgctag ccagccccga acccaccaaa aagcctcgca tcaacagctt
     2401 tgaggagcat gtggcctcca cctcggccgc cctgcccagc tgcctgcccc cggaggtgcc
     2461 cacgcagctg cctggacaag agttaaagag tggaattcct ggaagggacc ctgagccacc
     2521 agtccagccc cctgaagggc agagcattct ctggataccc agagaggtga ccgtggcatc
     2581 caggagtgga cccggggctg caccaagcat cccggagccc agactggtct cgcttcctca
     2641 cctttctggc tctgcggcaa gcggctccgg tggaggtgga tcaccggtcg ccaccatggt
     2701 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2761 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2821 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2881 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2941 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3001 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3061 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3121 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3181 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3241 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3301 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3361 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     3421 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3481 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3541 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3601 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3661 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3721 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3781 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3841 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3901 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3961 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4021 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4081 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4141 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4201 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4261 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4321 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4381 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4441 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4501 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4561 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4621 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4681 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4741 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4801 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4861 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4921 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4981 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5041 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5101 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5161 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5221 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5281 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5341 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5401 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5461 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5521 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5641 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5701 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5761 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5821 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5881 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5941 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6001 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6061 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6121 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6541 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6841 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6901 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6961 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7021 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7081 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7141 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7201 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7261 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7321 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7381 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7441 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7501 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7561 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7621 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7681 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7741 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7801 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7861 tgacgt
//
