LOCUS       V034259                 8025 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034259
VERSION     V034259
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             875..3538
                     /label="HMGCR(NM_000859)"
                     /note="HMGCR(NM_000859)"
                     /gene="HMGCR"
     polyA_signal    3592..3816
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3862..4290
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4304..4633
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4700..5494
                     /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    5668..5789
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5838..5854)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5862..5878
                     /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(5886..5916)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5931..5952
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6240..6825)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6996..7856)
                     /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(7857..7961)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagccgc caccatgttg tcaagacttt ttcgaatgca
      901 tggcctcttt gtggcctccc atccctggga agtcatagtg gggacagtga cactgaccat
      961 ctgcatgatg tccatgaaca tgtttactgg taacaataag atctgtggtt ggaattatga
     1021 atgtccaaag tttgaagagg atgttttgag cagtgacatt ataattctga caataacacg
     1081 atgcatagcc atcctgtata tttacttcca gttccagaat ttacgtcaac ttggatcaaa
     1141 atatattttg ggtattgctg gccttttcac aattttctca agttttgtat tcagtacagt
     1201 tgtcattcac ttcttagaca aagaattgac aggcttgaat gaagctttgc cctttttcct
     1261 acttttgatt gacctttcca gagcaagcac attagcaaag tttgccctca gttccaactc
     1321 acaggatgaa gtaagggaaa atattgctcg tggaatggca attttaggtc ctacgtttac
     1381 cctcgatgct cttgttgaat gtcttgtgat tggagttggt accatgtcag gggtacgtca
     1441 gcttgaaatt atgtgctgct ttggctgcat gtcagttctt gccaactact tcgtgttcat
     1501 gactttcttc ccagcttgtg tgtccttggt attagagctt tctcgggaaa gccgcgaggg
     1561 tcgtccaatt tggcagctca gccattttgc ccgagtttta gaagaagaag aaaataagcc
     1621 gaatcctgta actcagaggg tcaagatgat tatgtctcta ggcttggttc ttgttcatgc
     1681 tcacagtcgc tggatagctg atccttctcc tcaaaacagt acagcagata cttctaaggt
     1741 ttcattagga ctggatgaaa atgtgtccaa gagaattgaa ccaagtgttt ccctctggca
     1801 gttttatctc tctaaaatga tcagcatgga tattgaacaa gttattaccc taagtttagc
     1861 tctccttctg gctgtcaagt acatcttctt tgaacaaaca gagacagaat ctacactctc
     1921 attaaaaaac cctatcacat ctcctgtagt gacacaaaag aaagtcccag acaattgttg
     1981 tagacgtgaa cctatgctgg tcagaaataa ccagaaatgt gattcagtag aggaagagac
     2041 agggataaac cgagaaagaa aagttgaggt tataaaaccc ttagtggctg aaacagatac
     2101 cccaaacaga gctacatttg tggttggtaa ctcctcctta ctcgatactt catcagtact
     2161 ggtgacacag gaacctgaaa ttgaacttcc cagggaacct cggcctaatg aagaatgtct
     2221 acagatactt gggaatgcag agaaaggtgc aaaattcctt agtgatgctg agatcatcca
     2281 gttagtcaat gctaagcata tcccagccta caagttggaa actctgatgg aaactcatga
     2341 gcgtggtgta tctattcgcc gacagttact ttccaagaag ctttcagaac cttcttctct
     2401 ccagtaccta ccttacaggg attataatta ctccttggtg atgggagctt gttgtgagaa
     2461 tgttattgga tatatgccca tccctgttgg agtggcagga cccctttgct tagatgaaaa
     2521 agaatttcag gttccaatgg caacaacaga aggttgtctt gtggccagca ccaatagagg
     2581 ctgcagagca ataggtcttg gtggaggtgc cagcagccga gtccttgcag atgggatgac
     2641 tcgtggccca gttgtgcgtc ttccacgtgc ttgtgactct gcagaagtga aagcctggct
     2701 cgaaacatct gaagggttcg cagtgataaa ggaggcattt gacagcacta gcagatttgc
     2761 acgtctacag aaacttcata caagtatagc tggacgcaac ctttatatcc gtttccagtc
     2821 caggtcaggg gatgccatgg ggatgaacat gatttcaaag ggtacagaga aagcactttc
     2881 aaaacttcac gagtatttcc ctgaaatgca gattctagcc gttagtggta actattgtac
     2941 tgacaagaaa cctgctgcta taaattggat agagggaaga ggaaaatctg ttgtttgtga
     3001 agctgtcatt ccagccaagg ttgtcagaga agtattaaag actaccacag aggctatgat
     3061 tgaggtcaac attaacaaga atttagtggg ctctgccatg gctgggagca taggaggcta
     3121 caacgcccat gcagcaaaca ttgtcaccgc catctacatt gcctgtggac aggatgcagc
     3181 acagaatgtt ggtagttcaa actgtattac tttaatggaa gcaagtggtc ccacaaatga
     3241 agatttatat atcagctgca ccatgccatc tatagagata ggaacggtgg gtggtgggac
     3301 caacctacta cctcagcaag cctgtttgca gatgctaggt gttcaaggag catgcaaaga
     3361 taatcctggg gaaaatgccc ggcagcttgc ccgaattgtg tgtgggaccg taatggctgg
     3421 ggaattgtca cttatggcag cattggcagc aggacatctt gtcaaaagtc acatgattca
     3481 caacaggtcg aagatcaatt tacaagacct ccaaggagct tgcaccaaga agacagcctg
     3541 agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     3601 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3661 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3721 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3781 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3841 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3901 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3961 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4021 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4081 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4141 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4201 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4261 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4321 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4381 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4441 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4501 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4561 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4621 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4681 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4741 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4801 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4861 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4921 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4981 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5041 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5101 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5161 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5221 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5281 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5341 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5401 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5461 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5521 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5581 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5641 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5701 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5761 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5821 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5881 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5941 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6001 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6061 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6121 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6421 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6661 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt
     6781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7981 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
