LOCUS       V018428                 7686 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018428
VERSION     V018428
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             613..1329
                     /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             1369..4320
                     /label="PKN2(NM_006256)"
                     /note="PKN2(NM_006256)"
                     /gene="PKN2"
     polyA_signal    4474..4595
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4602..5057)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5084..5188
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        5190..5547
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5582..6376
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6608..6655
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6984..7572
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agctcaagct tcgaattcat ggcgtccaac
     1381 cccgaacggg gggagattct gctcacggaa ctgcaggggg attcccgaag tcttccgttt
     1441 tctgagaatg tgagtgctgt tcaaaaatta gacttttcag atacaatggt gcagcagaaa
     1501 ttggatgata tcaaggatcg aattaagaga gaaataagga aagaactgaa aatcaaagaa
     1561 ggagctgaaa atctgaggaa agtcacaaca gataaaaaaa gtttggctta tgtagacaac
     1621 attttgaaaa aatcaaataa aaaattagaa gaactacatc acaagctgca ggaattaaat
     1681 gcacatattg ttgtatcaga tccagaagat attacagatt gcccaaggac tccagatact
     1741 ccaaataatg accctcgttg ttctactagc aacaatagat tgaaggcctt acaaaaacaa
     1801 ttggatatag aacttaaagt aaaacaaggt gcagagaata tgatacagat gtattcaaat
     1861 ggatcttcaa aggatcggaa actccatggt acagctcagc aactgctcca ggacagcaag
     1921 acaaaaatag aagtcatacg aatgcagatt cttcaggcag tccagactaa tgaattggct
     1981 tttgataatg caaaacctgt gataagtcct cttgaacttc ggatggaaga attaaggcat
     2041 cattttagga tagagtttgc agtagcagaa ggtgcaaaga atgtaatgaa attacttggc
     2101 tcaggaaaag taacagacag aaaagcactt tcagaagctc aagcaagatt taatgaatca
     2161 agtcagaagt tggacctttt aaagtattca ttagagcaaa gattaaacga agtccccaag
     2221 aatcatccca aaagcaggat tattattgaa gaactttcac ttgttgctgc atcaccaaca
     2281 ctaagtccac gtcaaagtat gatatctacg caaaatcaat atagtacact atccaaacca
     2341 gcagcactaa caggtacttt ggaagttcgt cttatgggct gccaagatat cctagagaat
     2401 gtccctggac ggtcaaaagc aacatcagtt gcactgcctg gttggagtcc aagtgaaacc
     2461 agatcatctt tcatgagcag aacgagtaaa agtaaaagcg gaagtagtcg aaatcttcta
     2521 aaaaccgatg acttgtccaa tgatgtctgt gctgttttga agctcgataa tactgtggtt
     2581 ggccaaacta gctggaaacc catttccaat cagtcatggg accagaagtt tacactggaa
     2641 ctggacaggt cacgtgaact ggaaatttca gtttattggc gtgattggcg gtctctgtgt
     2701 gctgtaaaat ttctgaggtt agaagatttt ttagacaacc aacggcatgg catgtgtctc
     2761 tatttggaac cacagggtac tttatttgca gaggttacct tttttaatcc agttattgaa
     2821 agaagaccaa aacttcaaag acaaaagaaa attttttcaa agcaacaagg caaaacattt
     2881 ctcagagctc ctcaaatgaa tattaatatt gccacttggg gaaggctagt aagaagagct
     2941 attcctacag taaatcattc tggcaccttc agccctcaag ctcctgtgcc tactacagtg
     3001 ccagtggttg atgtacgcat ccctcaacta gcacctccag ctagtgattc tacagtaacc
     3061 aaattggact ttgatcttga gcctgaacct cctccagccc caccacgagc ttcttctctt
     3121 ggagaaatag atgaatcttc tgaattaaga gttttggata taccaggaca ggattcagag
     3181 actgtttttg atattcagaa tgacagaaat agtatacttc caaaatctca atctgaatac
     3241 aagcctgata ctcctcagtc aggcctagaa tatagtggta ttcaagaact tgaggacaga
     3301 agatctcagc aaaggtttca gtttaatcta caagatttca ggtgttgtgc tgtcttggga
     3361 agaggacatt ttggaaaggt gcttttagct gaatataaaa acacaaatga gatgtttgct
     3421 ataaaagcct taaagaaagg agatattgtg gctcgagatg aagtagacag cctgatgtgt
     3481 gaaaaaagaa tttttgaaac tgtgaatagt gtaaggcatc cctttttggt gaaccttttt
     3541 gcatgtttcc aaaccaaaga gcatgtttgc tttgtaatgg aatatgctgc cggtggggac
     3601 ctaatgatgc acattcatac tgatgtcttt tctgaaccaa gagctgtatt ttatgctgct
     3661 tgtgtagttc ttgggttgca gtatttacat gaacacaaaa ttgtttatag agatttgaaa
     3721 ttggataact tattgctaga tacagagggc tttgtgaaaa ttgctgattt tggtctttgc
     3781 aaagaaggaa tgggatatgg agatagaaca agcacatttt gtggcactcc tgaatttctt
     3841 gccccagaag tattaacaga aacttcttat acaagggctg tagattggtg gggccttggc
     3901 gtgcttatat atgaaatgct tgttggtgag tctccctttc ctggtgatga tgaagaggaa
     3961 gtttttgaca gtattgtaaa tgatgaagta aggtatccaa ggttcttatc tacagaagcc
     4021 atttctataa tgagaaggct gttaagaaga aatcctgaac ggcgccttgg ggctagcgag
     4081 aaagatgcag aggatgtaaa aaagcaccca tttttccggc taattgattg gagcgctctg
     4141 atggacaaaa aagtaaagcc accatttata cctaccataa gaggacgaga agatgttagt
     4201 aattttgatg atgaatttac ctcagaagca cctattctga ctccacctcg agaaccaagg
     4261 atactttcgg aagaggagca ggaaatgttc agagattttg actacattgc tgattggtgt
     4321 taagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     4381 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4441 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4501 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4561 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     4621 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4681 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4741 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4801 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4861 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4921 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4981 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5041 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5101 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5161 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     5221 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5281 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5341 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5401 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5461 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5521 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     5581 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5641 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5701 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5761 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5821 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5881 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5941 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     6001 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     6061 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     6121 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     6181 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     6241 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6301 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6361 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6421 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6481 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6541 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     6601 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6661 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     6721 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6781 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6841 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6901 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6961 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7021 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7081 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     7141 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7201 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7261 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7321 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7381 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7441 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7501 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7561 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7621 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7681 atgcat
//
