LOCUS       V026228                 9830 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026228
VERSION     V026228
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             902..1618
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             1634..4402
                     /label="RAPGEF3(NM_001098531)"
                     /note="RAPGEF3(NM_001098531)"
                     /gene="RAPGEF3"
     CDS             4418..5353
                     /codon_start=1
                     /note="human codon-optimized"
                     /product="Renilla luciferase"
                     /protein_id=""
                     /transl_table=1
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ*"
                     /label="hRluc"
     polyA_signal    5397..5621
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5667..6095
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6109..6438
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6505..7299
                     /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    7473..7594
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7643..7659)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7667..7683
                     /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(7691..7721)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7736..7757
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8045..8630)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8801..9661)
                     /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(9662..9766)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      961 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1081 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     1141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1321 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1381 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1501 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     1621 tggaggtgga tctatgaagg tgggctggcc aggtgagagc tgctggcagg tgggcctggc
     1681 tgtggaggat agcccagctc tgggagcacc gcgggtggga gccctccctg acgtggtgcc
     1741 ggaggggaca ctactcaaca tggtgttgag aaggatgcac cggccccgaa gctgctccta
     1801 ccagctgctg ctggagcacc agcgtccgag ctgcatccag gggctgcgct ggacaccact
     1861 caccaacagc gaggagtccc tggatttcag cgagagcctg gagcaggcct ccacagagcg
     1921 ggtgctcagg gctgggaggc agctgcatcg gcatctgctg gccacctgcc caaacctcat
     1981 ccgagaccgg aagtaccacc ttaggctcta tcggcagtgc tgctctggcc gggagctggt
     2041 ggatgggatc ttggccctgg gacttggggt ccattcccgg agccaagttg tgggaatctg
     2101 ccaggtgctg ctggatgaag gtgccctctg ccatgtgaaa cacgactggg ccttccagga
     2161 ccgagatgcc caattctacc ggttccccgg gcccgagccc gagcccgtga gaactcatga
     2221 gatggaggag gagttggccg aagctgtggc cctgctctcc cagcgggggc ctgacgccct
     2281 gctcactgtg gcacttcgaa agcccccagg tcagcgcacg gatgaagagc tggacctcat
     2341 ctttgaggag ctgctgcaca tcaaggctgt ggcccacctc tccaactcgg tgaagcgaga
     2401 attagcggct gttctgctct ttgaaccaca cagcaaggca gggaccgtgt tgttcagcca
     2461 gggggacaag ggcacttcgt ggtacattat ctggaaggga tctgtcaacg tggtgaccca
     2521 tggcaagggg ctggtgacca ccctgcatga gggagatgat tttggacagc tggctctggt
     2581 gaatgatgca ccccgggcag ccaccatcat cctgcgagaa gacaactgtc atttcctgcg
     2641 tgtggacaag caggacttca accgtatcat caaggatgtg gaggcaaaga ccatgcggct
     2701 ggaagaacat ggcaaagtgg tgctggtgct ggagagagcc tctcagggcg ccggcccttc
     2761 ccgaccccca accccaggca ggaaccggta tacagtgatg tctggcaccc cagagaagat
     2821 cctagagctt ctgttggagg ccatgggacc agattccagt gctcatgacc caacagagac
     2881 attcctcagc gacttcctcc tgacccacag ggtcttcatg cccagcgccc aactctgcgc
     2941 tgcccttctg caccacttcc atgtggagcc tgcgggtggc agcgagcagg agcgcagcac
     3001 ctacgtctgc aacaagaggc agcagatctt gcggctggtc agccagtggg tggccctgta
     3061 tggctccatg ctccacactg accctgtggc caccagcttc ctccagaaac tctcagacct
     3121 ggtgggcagg gacacccgac tcagcaacct gctgagggag cagtggccag agaggcggcg
     3181 atgccacagg ttggagaatg gctgtgggaa tgcatctcct cagatgaagg cccggaactt
     3241 gcctgtttgg ctccccaacc aggacgagcc ccttcctggc agcagctgtg ccatccaagt
     3301 tggggataaa gtcccctatg acatctgccg gccagaccac tcagtgttga ccctgcagct
     3361 gcctgtgaca gcctccgtga gagaggtgat ggcagcgttg gcccaggagg atggctggac
     3421 caaggggcag gtgctggtga aggtcaattc tgcaggtgat gccattggcc tgcagccaga
     3481 tgcccgtggt gtggccacat ctctggggct caatgagcgt ctctttgttg tcaacccaca
     3541 ggaagtgcat gagctgatcc cacaccctga ccagctgggg cccactgtgg gctctgctga
     3601 ggggctggac ctggtgagtg ccaaggacct ggcaggccag ctgacggacc acgactggag
     3661 cctcttcaac agtatccacc aggtggagct gatccactat gtgctgggcc cccagcatct
     3721 gcgggatgtc accaccgcca acctggagcg cttcatgcgc cgcttcaatg agctgcagta
     3781 ctgggtggcc accgagctgt gtctctgccc cgtgcccggc ccccgggccc agctgctcag
     3841 gaagttcatt aagctggcgg cccacctcaa ggagcagaag aatctcaatt ccttctttgc
     3901 cgtcatgttt ggcctcagca actcggccat cagccgccta gcccacacct gggagcggct
     3961 gcctcacaaa gtccggaagc tgtactccgc cctcgagagg ctgctggatc cctcatggaa
     4021 ccaccgggta taccgactgg ccctcgccaa gctctcccct cctgtcatcc ccttcatgcc
     4081 ccttcttctc aaagacatga ccttcattca tgagggaaac cacacactag tggagaatct
     4141 catcaacttt gagaagatga gaatgatggc cagagccgcg cggatgctgc accactgccg
     4201 aagccacaac cctgtgcctc tctcaccact cagaagccga gtttcccacc tccacgagga
     4261 cagccaggtg gcgaggattt ccacatgctc ggagcagtcc ctgagcaccc ggagtccagc
     4321 cagcacctgg gcttatgtcc agcagctgaa ggtcattgac aaccagcggg aactctcccg
     4381 cctctcccga gagctggagc caggaggtgg cggatcaatg gcttccaagg tgtacgaccc
     4441 cgagcaacgc aaacgcatga tcactgggcc tcagtggtgg gctcgctgca agcaaatgaa
     4501 cgtgctggac tccttcatca actactatga ttccgagaag cacgccgaga acgccgtgat
     4561 ttttctgcat ggtaacgctg cctccagcta cctgtggagg cacgtcgtgc ctcacatcga
     4621 gcccgtggct agatgcatca tccctgatct gatcggaatg ggtaagtccg gcaagagcgg
     4681 gaatggctca tatcgcctcc tggatcacta caagtacctc accgcttggt tcgagctgct
     4741 gaaccttcca aagaaaatca tctttgtggg ccacgactgg ggggcttgtc tggcctttca
     4801 ctactcctac gagcaccaag acaagatcaa ggccatcgtc catgctgaga gtgtcgtgga
     4861 cgtgatcgag tcctgggacg agtggcctga catcgaggag gatatcgccc tgatcaagag
     4921 cgaagagggc gagaaaatgg tgcttgagaa taacttcttc gtcgagacca tgctcccaag
     4981 caagatcatg cggaaactgg agcctgagga gttcgctgcc tacctggagc cattcaagga
     5041 gaagggcgag gttagacggc ctaccctctc ctggcctcgc gagatccctc tcgttaaggg
     5101 aggcaagccc gacgtcgtcc agattgtccg caactacaac gcctaccttc gggccagcga
     5161 cgatctgcct aagatgttca tcgagtccga ccctgggttc ttttccaacg ctattgtcga
     5221 gggagctaag aagttcccta acaccgagtt cgtgaaggtg aagggcctcc acttcagcca
     5281 ggaggacgct ccagatgaaa tgggtaagta catcaagagc ttcgtggagc gcgtgctgaa
     5341 gaacgagcag tagctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     5401 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5461 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5521 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5581 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     5641 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     5701 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5761 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5821 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5881 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5941 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6001 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6061 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     6121 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6181 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6241 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6301 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6361 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6421 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     6481 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     6541 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6601 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6661 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     6721 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6781 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6841 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6901 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6961 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     7021 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     7081 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     7141 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     7201 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     7261 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     7321 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     7381 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     7441 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     7501 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7561 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     7621 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7681 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7741 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7801 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7861 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7921 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7981 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8041 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8101 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8161 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8221 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8281 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8341 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8401 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8461 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8521 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8581 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8641 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8701 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8761 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8821 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8881 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8941 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     9001 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     9061 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     9121 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     9181 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     9241 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     9301 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     9361 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     9421 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     9481 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9541 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9601 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9661 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9721 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9781 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
