LOCUS       V019772                 7190 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019772
VERSION     V019772
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             625..3063
                     /label="Kat2b(NM_020005)"
                     /note="Kat2b(NM_020005)"
                     /gene="Kat2b"
     CDS             3136..3855
                     /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    3978..4099
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4106..4561)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4588..4692
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4694..5051
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5086..5880
                     /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    6112..6159
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6488..7076
                     /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 ccggactcag atctcgaggc caccatggcc gaggctggcg gggccgggtc cccagcactg
      661 cctcccgcgc ccccgcacgg ttccccccgg accctggcca ccgctgccgg gagctctgct
      721 tcctgcgggc cagcgaccgc ggtggccgcg gcgggcacgg ccgagggacc gggaggaggc
      781 ggctcggccc ggatcgccgt gaagaaggcg cagttgcgct ctgctccgcg ggcgaagaag
      841 ctggagaaac tcggcgtgta ctccgcctgc aaggcagagg agtcctgtaa atgcaatggc
      901 tggaagaacc ctaacccctc tcctactcca ccaagaggag acctccagca gataattgtc
      961 agtttgacag aatcctgtcg aagctgtagc catgcccttg ctgctcacgt ttctcacttg
     1021 gagaatgtgt cagaggaaga gatggacaga ctcctgggaa ttgtgttgga tgtggagtac
     1081 ctcttcacct gcgtccacaa agaagaagat gcagatacca aacaagtgta cttctaccta
     1141 ttcaagctct tgagaaagtc aattttacaa agaggaaaac ctgtggttga aggctccttg
     1201 gagaagaagc cgccatttga gaagcccagt attgaacagg gtgtgaacaa cttcgtgcag
     1261 tacaagttta gtcacttgcc atcgaaagag aggcagacaa cgatcgagct ggccaagatg
     1321 tttctgaacc gcatcaacta ctggcatctg gaggctccat ctcagcggag actacggtct
     1381 cccaatgatg acatctctgg atacaaggaa aactacacaa ggtggttgtg ctactgcaat
     1441 gtaccgcagt tctgtgacag cttacctcgg tacgaaacca caaaggtgtt tgggagaaca
     1501 ttgcttcgct cggtcttcac catcatgaga cgacagctct tggaacaagc cagacaggaa
     1561 aaagacaaac tgcctcttga gaaacgcacg cttatcctca cacatttccc aaagtttctg
     1621 tccatgttgg aagaagaagt gtatagtcaa aattctccta tctgggatca ggattttctc
     1681 tcagcctctt ccagaaccag cccgctagga atccaaacag taatcagtcc tcctgttact
     1741 gggacagcat tgttcagttc aaattccact tcccatgagc agatcaatgg agggagaaca
     1801 agtcctggat gcagaggctc ttctgggctt gaagcaaacc cgggagaaaa gaggaaaatg
     1861 aacaactctc atgctcccga ggaggccaag agatctcgag tgatggggga tattcccgtg
     1921 gaattgatca atgaggtcat gtctaccatc acagaccctg cagggatgct tggaccagag
     1981 accaattttc tgtcagccca ttcggccaga gatgaggcgg cacggctgga agaacgcagg
     2041 ggtgtcattg aattccacgt ggtgggcaac tccctgaacc agaaaccaaa caagaagatc
     2101 ctgatgtggc tcgtgggcct ccagaatgtg ttttcccacc agctgcccag aatgcccaaa
     2161 gagtacatca cacggctcgt ctttgacccg aaacacaaaa cccttgcttt aattaaagat
     2221 ggccgtgtca ttggtggtat ctgtttccgg atgtttccat cccagggatt cacagagatt
     2281 gttttctgtg cagtaacctc aaatgaacaa gtcaagggct atggaaccca cctgatgaac
     2341 catctcaaag aataccacat aaagcacgag atcctcaact tcctcacata tgcagatgag
     2401 tatgccatcg gctatttcaa gaagcagggt ttctccaaag aaatcaaaat acctaaaacc
     2461 aaatatgttg gctacatcaa ggattatgaa ggggccactt tgatgggatg tgagctgaac
     2521 cctcagatcc catacacaga gttctctgtc atcattaaaa agcagaagga gatcattaaa
     2581 aagctgatag aaagaaaaca agcccagatt cgaaaagtct accctggact ttcgtgtttc
     2641 aaagatggag ttcggcagat tcctatagaa agcattcctg gaatcagaga gacaggctgg
     2701 aaaccaagtg gaaaagagaa aagtaaagag cccaaagacc ctgagcagct ttacagcacc
     2761 ctcaagaaca tcctgcagca ggtgaagaac catccaaatg cttggccttt catggaacca
     2821 gtgaagagaa cagaagctcc gggatattat gaagttataa ggttccccat ggatctgaaa
     2881 accatgagtg aacgcctcag gaacaggtac tatgtgtcta agaagttatt catggcggac
     2941 ttgcaacgag tgttcaccaa ctgcaaggag tacaaccctc ccgagagcga gtactacaaa
     3001 tgcgccagca tcctggagaa gttcttcttc agtaaaatta aggaagcagg gttgattgac
     3061 aagggtaccg gcggcggagg gagtggagga gggggaagcg gaggcggagg gagtcaggat
     3121 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3181 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3241 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3301 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3361 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3421 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3481 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3541 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3601 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3661 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3721 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     3781 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3841 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     3901 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3961 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4021 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4081 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4141 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4201 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4261 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4321 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4381 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4441 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4501 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4561 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4741 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4801 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4861 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4921 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4981 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5041 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5101 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5161 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5221 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5281 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5341 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5401 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5461 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5521 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5581 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5641 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5701 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5761 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5821 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5881 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5941 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6001 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6061 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6121 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6181 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6241 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6301 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6361 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6421 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6481 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6601 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7081 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7141 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
