LOCUS       V020000                 6210 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020000
VERSION     V020000
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             1363..2865
                     /label="KCNJ3(NM_002239)"
                     /note="KCNJ3(NM_002239)"
                     /gene="KCNJ3"
     polyA_signal    2998..3119
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3126..3581)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3608..3712
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        3714..4071
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4106..4900
                     /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    5132..5179
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5508..6096
                     /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 ccggactcag atctcgagga ggagggggaa gcatgtctgc actccgaagg
     1381 aaatttgggg acgattatca ggtagtgacc acatcgtcca gcggctcggg cttgcagccc
     1441 caggggccag gccaggaccc tcagcagcag cttgtgccca agaagaagcg gcagcggttc
     1501 gtggacaaga acggccggtg caatgtacag cacggcaacc tgggcagcga gacaagccgc
     1561 tacctctcgg acctcttcac cacgctggtg gacctcaagt ggcgctggaa cctcttcatc
     1621 ttcattctca cctacaccgt ggcctggctt ttcatggcgt ccatgtggtg ggtgatcgcc
     1681 tacactcggg gcgacctgaa caaagcccac gtcggtaact acacgccttg cgtggccaat
     1741 gtctataact tcccttctgc cttcctcttc ttcatcgaga cggaggccac catcggctat
     1801 ggctaccgat acatcacaga caagtgcccc gagggcatca tcctcttcct cttccagtcc
     1861 atcctgggct ccatcgtgga cgccttcctc atcggctgca tgttcatcaa gatgtcccag
     1921 cccaagaagc gcgccgagac cctcatgttc agcgagcacg cggtgatctc catgagggac
     1981 ggaaaactca cgcttatgtt ccgggtgggc aacctgcgca acagccacat ggtctccgcg
     2041 cagattcgct gcaagctgct caaatctcgg cagacacctg agggtgagtt ccttcccctt
     2101 gaccaacttg aactggatgt aggttttagt acaggggcag atcaactttt tcttgtgtcc
     2161 cccctcacaa tttgccacgt gatcgatgcc aaaagcccct tttatgacct atcccagcga
     2221 agcatgcaaa ctgaacagtt cgagattgtc gtcatcctag aaggcattgt ggaaacaact
     2281 gggatgactt gtcaagctcg aacatcatat actgaagatg aagttctttg gggtcatcgt
     2341 ttttttcctg taatttcctc agaagaggga ttctttaaag ttgattactc ccagttccat
     2401 gcaacatttg aagtccccac cccaccttac agtgtgaaag agcaggagga aatgcttctc
     2461 atgtcgtccc ctttaatagc accagccata actaacagca aagaaagaca taattctgtg
     2521 gaatgcttag atggactaga tgatattact acaaaactac catctaagct gcagaaaatt
     2581 actggaagag aagactttcc caaaaaactc ttgaggatga gttctacaac ttcagaaaaa
     2641 gcctacagct tgggagactt gcccatgaaa cttcaacgaa taagttcagt tccgggcaac
     2701 tcagaagaaa aactggtatc taaaaccacc aagatgttat ctgatcccat gagccagtct
     2761 gtggctgatt tgccaccaaa gcttcaaaag atggctggag gagcagctag gatggaaggg
     2821 aaccttccag ccaaattaag aaaaatgaac tctgatcgct tcacataagg atccaccgga
     2881 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2941 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3001 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3061 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3121 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3181 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3241 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3301 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3361 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3421 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3481 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3541 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3721 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3901 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3961 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4021 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4081 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4141 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4201 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4261 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4321 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4381 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4441 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4501 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4561 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4621 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4681 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4741 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4801 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4861 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4921 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4981 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5041 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5101 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5161 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5221 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5281 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5341 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5401 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5461 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5521 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5581 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5641 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5701 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5761 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5821 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5881 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5941 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6001 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6061 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6121 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6181 tctgtggata accgtattac cgccatgcat
//
