LOCUS       V042147                 7552 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042147
VERSION     V042147
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3042
                     /label="GMPS(NM_003875)"
                     /note="GMPS(NM_003875)"
                     /gene="GMPS"
     CDS             3049..3114
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3152..3376
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3422..3850
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3864..4193
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4260..5054
                     /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    5228..5349
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5398..5414)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5422..5438
                     /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(5446..5476)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5491..5512
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5800..6385)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6556..7416)
                     /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(7417..7521)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggctc tgtgcaacgg agactccaag ctggagaatg ctggaggaga ccttaaggat
     1021 ggccaccacc actatgaagg agctgttgtc attctggatg ctggtgctca gtacgggaaa
     1081 gtcatagacc gaagagtgag ggaactgttc gtgcagtctg aaattttccc cttggaaaca
     1141 ccagcatttg ctataaagga acaaggattc cgtgctatta tcatctctgg aggacctaat
     1201 tctgtgtatg ctgaagatgc tccctggttt gatccagcaa tattcactat tggcaagcct
     1261 gttcttggaa tttgctatgg tatgcagatg atgaataagg tatttggagg tactgtgcac
     1321 aaaaaaagtg tcagagaaga tggagttttc aacattagtg tggataatac atgttcatta
     1381 ttcaggggcc ttcagaagga agaagttgtt ttgcttacac atggagatag tgtagacaaa
     1441 gtagctgatg gattcaaggt tgtggcacgt tctggaaaca tagtagcagg catagcaaat
     1501 gaatctaaaa agttatatgg agcacagttc caccctgaag ttggccttac agaaaatgga
     1561 aaagtaatac tgaagaattt cctttatgat atagctggat gcagtggaac cttcaccgtg
     1621 cagaacagag aacttgagtg tattcgagag atcaaagaga gagtaggcac gtcaaaagtt
     1681 ttggttttac tcagtggtgg agtagactca acagtttgta cagctttgct aaatcgtgct
     1741 ttgaaccaag aacaagtcat tgctgtgcac attgataatg gctttatgag aaaacgagaa
     1801 agccagtctg ttgaagaggc cctcaaaaag cttggaattc aggtcaaagt gataaatgct
     1861 gctcattctt tctacaatgg aacaacaacc ctaccaatat cagatgaaga tagaacccca
     1921 cggaaaagaa ttagcaaaac gttaaatatg accacaagtc ctgaagagaa aagaaaaatc
     1981 attggggata cttttgttaa gattgccaat gaagtaattg gagaaatgaa cttgaaacca
     2041 gaggaggttt tccttgccca aggtacttta cggcctgatc taattgaaag tgcatccctt
     2101 gttgcaagtg gcaaagctga actcatcaaa acccatcaca atgacacaga gctcatcaga
     2161 aagttgagag aggagggaaa agtaatagaa cctctgaaag attttcataa agatgaagtg
     2221 agaattttgg gcagagaact tggacttcca gaagagttag tttccaggca tccatttcca
     2281 ggtcctggcc tggcaatcag agtaatatgt gctgaagaac cttatatttg taaggacttt
     2341 cctgaaacca acaatatttt gaaaatagta gctgattttt ctgcaagtgt taaaaagcca
     2401 cataccctat tacagagagt caaagcctgc acaacagaag aggatcagga gaagctgatg
     2461 caaattacca gtctgcattc actgaatgcc ttcttgctgc caattaaaac tgtaggtgtg
     2521 cagggtgact gtcgttccta cagttacgtg tgtggaatct ccagtaaaga tgaacctgac
     2581 tgggaatcac ttatttttct ggctaggctt atacctcgca tgtgtcacaa cgttaacaga
     2641 gttgtttata tatttggccc accagttaaa gaacctccta cagatgttac tcccactttc
     2701 ttgacaacag gggtgctcag tactttacgc caagctgatt ttgaggccca taacattctc
     2761 agggagtctg ggtatgctgg gaaaatcagc cagatgccgg tgattttgac accattacat
     2821 tttgatcggg acccacttca aaagcagcct tcatgccaga gatctgtggt tattcgaacc
     2881 tttattacta gtgacttcat gactggtata cctgcaacac ctggcaatga gatccctgta
     2941 gaggtggtat taaagatggt cactgagatt aagaagattc ctggtatttc tcgaattatg
     3001 tatgacttaa catcaaagcc cccaggaact actgagtggg agctcgagga ctacaaagac
     3061 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga
     3121 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3181 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3241 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3301 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3361 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3421 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3481 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3541 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3601 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3661 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3721 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3781 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3841 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3901 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4021 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4081 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4141 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4201 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4261 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4321 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4381 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4441 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4501 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4561 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4621 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4681 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4741 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4801 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4861 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4921 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4981 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5041 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5101 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5161 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5221 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5281 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5341 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5401 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5461 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5521 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5581 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5641 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5701 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5761 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5821 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5881 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5941 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6001 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6061 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6121 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6181 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6241 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6301 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6361 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6421 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6481 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6541 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6601 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6661 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6721 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6781 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6841 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6901 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6961 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7021 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7081 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7141 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7201 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7261 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7321 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7381 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7441 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7501 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
