LOCUS       V042291                 7588 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042291
VERSION     V042291
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             648..1301
                     /codon_start=1
                     /gene=""
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /transl_table=1
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
                     /label="GST"
     CDS             1338..1523
                     /label="Pcp4(NM_008791)"
                     /note="Pcp4(NM_008791)"
                     /gene="Pcp4"
     CDS             1560..2279
                     /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"
     misc_feature    2364..2952
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3018..3066
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3268..3696
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3710..4039
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5076..5197
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5246..5262)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5270..5286
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5294..5324)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5339..5360
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5648..6236)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6407..7267)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7268..7372)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcccaccatg tcccctatac
      661 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
      721 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
      781 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
      841 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
      901 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
      961 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
     1021 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     1081 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     1141 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     1201 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     1261 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa aggaggaggg ggaagcggcg
     1321 gcggagggag taagcttatg agtgagagac aaagtgccgg agcgaccaac ggaaaagaca
     1381 agacgtcagg agataatgat gggcagaaga aagtccaaga agaatttgat atcgacatgg
     1441 atgcaccaga gacagagcgt gcagctgtgg ccattcagtc tcagttcaga aaattccaga
     1501 agaaaaaggc aggatcacag tccggtaccg gcggtggagg aagcggcgga ggtggatcta
     1561 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1621 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1681 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1741 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1801 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1861 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1921 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1981 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2041 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2101 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2161 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2221 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtagg
     2281 atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag
     2341 tctagagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     2401 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2461 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2521 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2581 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2641 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2701 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     2761 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     2821 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2881 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2941 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     3001 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     3061 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     3121 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     3181 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     3241 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3301 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3361 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3421 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3481 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3541 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3601 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     3661 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3721 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3781 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3841 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3901 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3961 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4021 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4081 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4141 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4201 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4261 cctgtccggt gccctgaatg aactgcagga 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 gcgcgcatgc 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 ggggttcgcg
     4921 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4981 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5041 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5101 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5161 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5221 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5281 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5341 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5401 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5461 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5521 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5581 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5641 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5701 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5761 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5821 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     5881 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5941 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6001 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6061 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6121 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6181 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6241 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6301 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6361 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6421 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6481 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6541 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6601 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6661 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6721 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6781 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6841 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6901 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6961 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7021 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7081 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7141 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7201 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7261 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7321 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7381 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     7441 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     7501 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     7561 cgacaattgc atgaagaatc tgcttagg
//
