LOCUS       V025834                 6899 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025834
VERSION     V025834
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             624..2216
                     /label="CYP4F22(NM_173483)"
                     /note="CYP4F22(NM_173483)"
                     /gene="CYP4F22"
     misc_feature    2258..2844
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             2845..3564
                     /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    3687..3808
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3815..4270)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4297..4401
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4403..4760
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4795..5589
                     /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    5821..5868
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6197..6785
                     /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 atctcgagcc accatgctgc ccatcacaga ccgcctgctg cacctcctgg
      661 ggctggagaa gacggcgttc cgcatatacg cggtgtccac ccttctcctc ttcctgctct
      721 tcttcctgtt ccgcctgctg ctgcggttcc tgaggctctg caggagcttc tacatcacct
      781 gccgccggct gcgctgcttc ccccagcctc cccggcgcaa ctggctgctg ggccacctgg
      841 gcatgtacct tccaaatgag gcgggccttc aagatgagaa gaaggtactg gacaacatgc
      901 accatgtact cttggtatgg atgggacctg tcctgccgct gttggttctg gtgcaccctg
      961 attacatcaa accccttttg ggagcctcag ctgccatcgc ccccaaggat gacctcttct
     1021 atggcttcct aaaaccttgg ctaggggatg ggctgctgct cagcaaaggt gacaagtgga
     1081 gccggcaccg tcgcctgctg acacccgcct tccactttga catcctgaag ccttacatga
     1141 agatcttcaa ccagagcgct gacattatgc atgctaaatg gcggcatctg gcagagggct
     1201 cagcggtctc ccttgatatg tttgagcata tcagcctcat gaccctggac agtcttcaga
     1261 aatgtgtctt cagctacaac agcaactgcc aagagaagat gagtgattat atctccgcta
     1321 tcattgaact gagcgctctg tctgtccggc gccagtatcg cttgcaccac tacctcgact
     1381 tcatttacta ccgctcggcg gatgggcgga ggttccggca ggcctgtgac atggtgcacc
     1441 acttcaccac tgaagtcatc caggaacggc ggcgggcact gcgtcagcag ggggccgagg
     1501 cctggcttaa ggccaagcag gggaagacct tggactttat tgatgtgctg ctcctggcca
     1561 gggatgaaga tggaaaggaa ctgtcagacg aggatatccg agccgaagca gacaccttca
     1621 tgtttgaggg tcacgacaca acatccagtg ggatctcttg gatgctgttc aatttggcaa
     1681 agtatccgga ataccaggag aaatgccgag aagagattca ggaagtcatg aaaggccggg
     1741 agctggagga gctggagtgg gacgatctga ctcagctgcc ctttacaact atgtgcatta
     1801 aggagagcct gcgccagtac ccacctgtca ctcttgtctc tcgccaatgc acggaggaca
     1861 tcaagctccc agatgggcgc atcatcccca aaggaatcat ctgcttggtc agcatctatg
     1921 gaacccacca caaccccaca gtgtggcctg actccaaggt gtacaacccc taccgctttg
     1981 acccggacaa cccacagcag cgctctccac tggcctatgt gcccttctct gcaggaccca
     2041 ggaattgcat cggacagagc ttcgccatgg ccgagttgcg cgtggttgtg gcactaacac
     2101 tgctacgttt ccgcctgagc gtggaccgaa cgcgcaaggt gcggcggaag ccggagctca
     2161 tactgcgcac ggagaacggg ctctggctca aggtggagcc gctgcctccg cgggcctgag
     2221 aattctgcag tcgacggtac cgcgggcccg ggatccgccc ctctccctcc ccccccccta
     2281 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     2341 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     2401 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     2461 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     2521 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     2581 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     2641 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     2701 taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt
     2761 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     2821 cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg
     2881 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2941 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3001 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3061 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3121 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3181 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3241 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3301 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3361 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3421 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3481 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3541 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     3601 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3661 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3721 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3781 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3841 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3901 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3961 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4021 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4081 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4141 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4201 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4261 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4321 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4381 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4441 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4501 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4561 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4621 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4681 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4741 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4801 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4861 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4921 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4981 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5041 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5101 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5161 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5221 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5281 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5341 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5401 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5461 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5521 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5581 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5641 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5701 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5761 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5821 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5881 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5941 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6001 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6061 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6781 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6841 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
