LOCUS       V038191                 9236 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038191
VERSION     V038191
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             908..1624
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced CFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNG
                     IKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="ECFP"
     CDS             1631..4756
                     /label="ATP2A2(NM_170665)"
                     /note="ATP2A2(NM_170665)"
                     /gene="ATP2A2"
     polyA_signal    4803..5027
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5073..5501
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5515..5844
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5911..6705
                     /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    6879..7000
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7049..7065)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7073..7089
                     /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(7097..7127)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7142..7163
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7451..8036)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8207..9067)
                     /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(9068..9172)
                     /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 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1081 gcccaccctc gtgaccaccc tgacctgggg cgtgcagtgc ttcagccgct accccgacca
     1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1321 ggggcacaag ctggagtaca actacatcag ccacaacgtc tatatcaccg ccgacaagca
     1381 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1501 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1621 caaggaattc atggagaacg cgcacaccaa gacggtggag gaggtgctgg gccacttcgg
     1681 cgtcaacgag agtacggggc tgagcctgga acaggtcaag aagcttaagg agagatgggg
     1741 ctccaacgag ttaccggctg aagaaggaaa aaccttgctg gaacttgtga ttgagcagtt
     1801 tgaagacttg ctagttagga ttttattact ggcagcatgt atatcttttg ttttggcttg
     1861 gtttgaagaa ggtgaagaaa caattacagc ctttgtagaa ccttttgtaa ttttactcat
     1921 attagtagcc aatgcaattg tgggtgtatg gcaggaaaga aatgctgaaa atgccatcga
     1981 agcccttaag gaatatgagc ctgaaatggg caaagtgtat cgacaggaca gaaagagtgt
     2041 gcagcggatt aaagctaaag acatagttcc tggtgatatt gtagaaattg ctgttggtga
     2101 caaagttcct gctgatataa ggttaacttc catcaaatct accacactaa gagttgacca
     2161 gtcaattctc acaggtgaat ctgtctctgt catcaagcac actgatcccg tccctgaccc
     2221 acgagctgtc aaccaagata aaaagaacat gctgttttct ggtacaaaca ttgctgctgg
     2281 gaaagctatg ggagtggtgg tagcaactgg agttaacacc gaaattggca agatccggga
     2341 tgaaatggtg gcaacagaac aggagagaac accccttcag caaaaactag atgaatttgg
     2401 ggaacagctt tccaaagtca tctcccttat ttgcattgca gtctggatca taaatattgg
     2461 gcacttcaat gacccggttc atggagggtc ctggatcaga ggtgctattt actactttaa
     2521 aattgcagtg gccctggctg tagcagccat tcctgaaggt ctgcctgcag tcatcaccac
     2581 ctgcctggct cttggaactc gcagaatggc aaagaaaaat gccattgttc gaagcctccc
     2641 gtctgtggaa acccttggtt gtacttctgt tatctgctca gacaagactg gtacacttac
     2701 aacaaaccag atgtcagtct gcaggatgtt cattctggac agagtggaag gtgatacttg
     2761 ttcccttaat gagtttacca taactggatc aacttatgca cctattggag aagtgcataa
     2821 agatgataaa ccagtgaatt gtcaccagta tgatggtctg gtagaattag caacaatttg
     2881 tgctctttgt aatgactctg ctttggatta caatgaggca aagggtgtgt atgaaaaagt
     2941 tggagaagct acagagactg ctctcacttg cctagtagag aagatgaatg tatttgatac
     3001 cgaattgaag ggtctttcta aaatagaacg tgcaaatgcc tgcaactcag tcattaaaca
     3061 gctgatgaaa aaggaattca ctctagagtt ttcacgtgac agaaagtcaa tgtcggttta
     3121 ctgtacacca aataaaccaa gcaggacatc aatgagcaag atgtttgtga agggtgctcc
     3181 tgaaggtgtc attgacaggt gcacccacat tcgagttgga agtactaagg ttcctatgac
     3241 ctctggagtc aaacagaaga tcatgtctgt cattcgagag tggggtagtg gcagcgacac
     3301 actgcgatgc ctggccctgg ccactcatga caacccactg agaagagaag aaatgcacct
     3361 tgaggactct gccaacttta ttaaatatga gaccaatctg accttcgttg gctgcgtggg
     3421 catgctggat cctccgagaa tcgaggtggc ctcctccgtg aagctgtgcc ggcaagcagg
     3481 catccgggtc atcatgatca ctggggacaa caagggcact gctgtggcca tctgtcgccg
     3541 catcggcatc ttcgggcagg atgaggacgt gacgtcaaaa gctttcacag gccgggagtt
     3601 tgatgaactc aacccctccg cccagcgaga cgcctgcctg aacgcccgct gttttgctcg
     3661 agttgaaccc tcccacaagt ctaaaatcgt agaatttctt cagtcttttg atgagattac
     3721 agctatgact ggcgatggcg tgaacgatgc tcctgctctg aagaaagccg agattggcat
     3781 tgctatgggc tctggcactg cggtggctaa aaccgcctct gagatggtcc tggcggatga
     3841 caacttctcc accattgtgg ctgccgttga ggaggggcgg gcaatctaca acaacatgaa
     3901 acagttcatc cgctacctca tctcgtccaa cgtcggggaa gttgtctgta ttttcctgac
     3961 agcagccctt ggatttcccg aggctttgat tcctgttcag ctgctctggg tcaatctggt
     4021 gacagatggc ctgcctgcca ctgcactggg gttcaaccct cctgatctgg acatcatgaa
     4081 taaacctccc cggaacccaa aggaaccatt gatcagcggg tggctctttt tccgttactt
     4141 ggctattggc tgttacgtcg gcgctgctac cgtgggtgct gctgcatggt ggttcattgc
     4201 tgctgacggt ggtccaagag tgtccttcta ccagctgagt catttcctac agtgtaaaga
     4261 ggacaacccg gactttgaag gcgtggattg tgcaatcttt gaatccccat acccgatgac
     4321 aatggcgctc tctgttctag taactataga aatgtgtaac gccctcaaca gcttgtccga
     4381 aaaccagtcc ttgctgagga tgcccccctg ggagaacatc tggctcgtgg gctccatctg
     4441 cctgtccatg tcactccact tcctgatcct ctatgtcgaa cccttgccac tcatcttcca
     4501 gatcacaccg ctgaacgtga cccagtggct gatggtgctg aaaatctcct tgcccgtgat
     4561 tctcatggat gagacgctca agtttgtggc ccgcaactac ctggaacctg gtaaagagtg
     4621 tgtgcagcct gccaccaaat cctgctcgtt ctcggcatgc accgatggga tttcctggcc
     4681 gtttgtgctg ctcataatgc ccctggtgat ctgggtctat agcacagaca ctaactttag
     4741 cgatatgttc tggtcttgac tcgagtctag agggcccgtt taaacccgct gatcagcctc
     4801 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4861 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4921 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4981 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     5041 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     5101 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5161 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5221 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5281 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5341 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5401 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5461 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     5521 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5581 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5641 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5761 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     5821 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     5881 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5941 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     6001 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     6061 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     6121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6421 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6721 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6781 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6841 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6901 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6961 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     7021 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7081 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     7141 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7201 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7261 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7321 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7861 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7981 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9181 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
