LOCUS       V018333                 7945 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018333
VERSION     V018333
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             603..1079
                     /EC_number=""
                     /codon_start=1
                     /experiment=""
                     /function=""
                     /gene="folA"
                     /gene_synonym=""
                     /locus_tag=""
                     /note=""
                     /product="E. coli dihydrofolate reductase"
                     /protein_id=""
                     /transl_table=1
                     /translation="MISLIAALAVDHVIGMETVMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
                     /label="ecDHFR"
     CDS             1122..3857
                     /codon_start=1
                     /product="DNA methyltransferase 3A"
                     /transl_table=1
                     /translation="MPAMPSSGPGDTSSSAAEREEDRKDGEEQEEPRGKEERQEPSTTA
                     RKVGRPGRKRKHPPVESGDTPKDPAVISKSPSMAQDSGASELLPNGDLEKRSEPQPEEG
                     SPAGGQKGGAPAEGEGAAETLPEASRAVENGCCTPKEGRGAPAEAGKEQKETNIESMKM
                     EGSRGRLRGGLGWESSLRQRPMPRLTFQAGDPYYISKRKRDEWLARWKREAEKKAKVIA
                     GMNAVEENQGPGESQKVEEASPPAVQQPTDPASPTVATTPEPVGSDAGDKNATKAGDDE
                     PEYEDGRGFGIGELVWGKLRGFSWWPGRIVSWWMTGRSRAAEGTRWVMWFGDGKFSVVC
                     VEKLMPLSSFCSAFHQATYNKQPMYRKAIYEVLQVASSRAGKLFPVCHDSDESDTAKAV
                     EVQNKPMIEWALGGFQPSGPKGLEPPEEEKNPYKEVYTDMWVEPEAAAYAPPPPAKKPR
                     KSTAEKPKVKEIIDERTRERLVYEVRQKCRNIEDICISCGSLNVTLEHPLFVGGMCQNC
                     KNCFLECAYQYDDDGYQSYCTICCGGREVLMCGNNNCCRCFCVECVDLLVGPGAAQAAI
                     KEDPWNCYMCGHKGTYGLLRRREDWPSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIR
                     VLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKH
                     IQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFW
                     LFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDK
                     LELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFP
                     VHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV"
                     /label="DNMT3A"
     CDS             3891..4610
                     /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    4733..4854
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4861..5316)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5343..5447
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        5449..5806
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5841..6635
                     /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    6867..6914
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      7243..7831
                     /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 gctagcgcca
      601 ccatgatcag tctgattgcg gcgttagcgg tagatcacgt tatcggcatg gaaaccgtca
      661 tgccgtggaa cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg
      721 tgattatggg ccgccatacc tgggaatcaa tcggtcgtcc gttgccagga cgcaaaaata
      781 ttatcctcag cagtcaaccg agtacggacg atcgcgtaac gtgggtgaag tcggtggatg
      841 aagccatcgc ggcgtgtggt gacgtaccag aaatcatggt tattggcggc ggtcgcgttt
      901 atgaacagtt cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgac gcagaagtgg
      961 aaggcgacac ccatttcccg gattacgagc cggatgactg ggaatcggta ttcagcgaat
     1021 tccacgatgc tgatgcgcag aactctcaca gctattgctt tgagattctg gagcggcgag
     1081 gcggcggagg gagtggagga gggggaagca agcttgccac catgcccgcc atgccctcca
     1141 gcggccccgg ggacaccagc agctctgctg cggagcggga ggaggaccga aaggacggag
     1201 aggagcagga ggagccgcgt ggcaaggagg agcgccaaga gcccagcacc acggcacgga
     1261 aggtggggcg gcctgggagg aagcgcaagc accccccggt ggaaagcggt gacacgccaa
     1321 aggaccctgc ggtgatctcc aagtccccat ccatggccca ggactcaggc gcctcagagc
     1381 tattacccaa tggggacttg gagaagcgga gtgagcccca gccagaggag gggagccctg
     1441 ctggggggca gaagggcggg gccccagcag agggagaggg tgcagctgag accctgcctg
     1501 aagcctcaag agcagtggaa aatggctgct gcacccccaa ggagggccga ggagcccctg
     1561 cagaagcggg caaagaacag aaggagacca acatcgaatc catgaaaatg gagggctccc
     1621 ggggccggct gcggggtggc ttgggctggg agtccagcct ccgtcagcgg cccatgccga
     1681 ggctcacctt ccaggcgggg gacccctact acatcagcaa gcgcaagcgg gacgagtggc
     1741 tggcacgctg gaaaagggag gctgagaaga aagccaaggt cattgcagga atgaatgctg
     1801 tggaagaaaa ccaggggccc ggggagtctc agaaggtgga ggaggccagc cctcctgctg
     1861 tgcagcagcc cactgacccc gcatccccca ctgtggctac cacgcctgag cccgtggggt
     1921 ccgatgctgg ggacaagaat gccaccaaag caggcgatga cgagccagag tacgaggacg
     1981 gccggggctt tggcattggg gagctggtgt gggggaaact gcggggcttc tcctggtggc
     2041 caggccgcat tgtgtcttgg tggatgacgg gccggagccg agcagctgaa ggcacccgct
     2101 gggtcatgtg gttcggagac ggcaaattct cagtggtgtg tgttgagaag ctgatgccgc
     2161 tgagctcgtt ttgcagtgcg ttccaccagg ccacgtacaa caagcagccc atgtaccgca
     2221 aagccatcta cgaggtcctg caggtggcca gcagccgcgc ggggaagctg ttcccggtgt
     2281 gccacgacag cgatgagagt gacactgcca aggccgtgga ggtgcagaac aagcccatga
     2341 ttgaatgggc cctggggggc ttccagcctt ctggccctaa gggcctggag ccaccagaag
     2401 aagagaagaa tccctacaaa gaagtgtaca cggacatgtg ggtggaacct gaggcagctg
     2461 cctacgcacc acctccacca gccaaaaagc cccggaagag cacagcggag aagcccaagg
     2521 tcaaggagat tattgatgag cgcacaagag agcggctggt gtacgaggtg cggcagaagt
     2581 gccggaacat tgaggacatc tgcatctcct gtgggagcct caatgttacc ctggaacacc
     2641 ccctcttcgt tggaggaatg tgccaaaact gcaagaactg ctttctggag tgtgcgtacc
     2701 agtacgacga cgacggctac cagtcctact gcaccatctg ctgtgggggc cgtgaggtgc
     2761 tcatgtgcgg aaacaacaac tgctgcaggt gcttttgcgt ggagtgtgtg gacctcttgg
     2821 tggggccggg ggctgcccag gcagccatta aggaagaccc ctggaactgc tacatgtgcg
     2881 ggcacaaggg tacctacggg ctgctgcggc ggcgagagga ctggccctcc cggctccaga
     2941 tgttcttcgc taataaccac gaccaggaat ttgaccctcc aaaggtttac ccacctgtcc
     3001 cagctgagaa gaggaagccc atccgggtgc tgtctctctt tgatggaatc gctacagggc
     3061 tcctggtgct gaaggacttg ggcattcagg tggaccgcta cattgcctcg gaggtgtgtg
     3121 aggactccat cacggtgggc atggtgcggc accaggggaa gatcatgtac gtcggggacg
     3181 tccgcagcgt cacacagaag catatccagg agtggggccc attcgatctg gtgattgggg
     3241 gcagtccctg caatgacctc tccatcgtca accctgctcg caagggcctc tacgagggca
     3301 ctggccggct cttctttgag ttctaccgcc tcctgcatga tgcgcggccc aaggagggag
     3361 atgatcgccc cttcttctgg ctctttgaga atgtggtggc catgggcgtt agtgacaaga
     3421 gggacatctc gcgatttctc gagtccaacc ctgtgatgat tgatgccaaa gaagtgtcag
     3481 ctgcacacag ggcccgctac ttctggggta accttcccgg tatgaacagg ccgttggcat
     3541 ccactgtgaa tgataagctg gagctgcagg agtgtctgga gcatggcagg atagccaagt
     3601 tcagcaaagt gaggaccatt actacgaggt caaactccat aaagcagggc aaagaccagc
     3661 attttcctgt cttcatgaat gagaaagagg acatcttatg gtgcactgaa atggaaaggg
     3721 tatttggttt cccagtccac tatactgacg tctccaacat gagccgcttg gcgaggcaga
     3781 gactgctggg ccggtcatgg agcgtgccag tcatccgcca cctcttcgct ccgctgaagg
     3841 agtattttgc gtgtgtgtgc ccgcgggccc gggatccacc ggtcgccacc atggtgagca
     3901 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3961 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     4021 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4081 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     4141 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4201 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4261 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4321 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     4381 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     4441 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     4501 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     4561 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg
     4621 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     4681 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     4741 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4801 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     4861 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4921 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4981 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5041 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5101 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5161 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5221 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5281 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     5341 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5401 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     5461 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5521 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5581 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5641 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5701 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5761 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     5821 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5881 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5941 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     6001 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     6061 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6121 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6181 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6241 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6301 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6361 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6421 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6481 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6541 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6601 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6661 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6721 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6781 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     6841 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6901 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6961 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     7021 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     7081 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7141 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7201 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7261 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7321 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7381 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7441 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7501 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7561 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7621 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7681 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7741 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7801 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7861 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7921 ggataaccgt attaccgcca tgcat
//
