LOCUS       V035089                 8791 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035089
VERSION     V035089
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..4281
                     /label="PDGFRB(NM_002609)"
                     /note="PDGFRB(NM_002609)"
                     /gene="PDGFRB"
     CDS             4288..4353
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4391..4615
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4661..5089
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5103..5432
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5499..6293
                     /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    6467..6588
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6637..6653)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6661..6677
                     /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(6685..6715)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6730..6751
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7039..7624)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7795..8655)
                     /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(8656..8760)
                     /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 accatgcggc ttccgggtgc gatgccagct ctggccctca aaggcgagct gctgttgctg
     1021 tctctcctgt tacttctgga accacagatc tctcagggcc tggtcgtcac acccccgggg
     1081 ccagagcttg tcctcaatgt ctccagcacc ttcgttctga cctgctcggg ttcagctccg
     1141 gtggtgtggg aacggatgtc ccaggagccc ccacaggaaa tggccaaggc ccaggatggc
     1201 accttctcca gcgtgctcac actgaccaac ctcactgggc tagacacggg agaatacttt
     1261 tgcacccaca atgactcccg tggactggag accgatgagc ggaaacggct ctacatcttt
     1321 gtgccagatc ccaccgtggg cttcctccct aatgatgccg aggaactatt catctttctc
     1381 acggaaataa ctgagatcac cattccatgc cgagtaacag acccacagct ggtggtgaca
     1441 ctgcacgaga agaaagggga cgttgcactg cctgtcccct atgatcacca acgtggcttt
     1501 tctggtatct ttgaggacag aagctacatc tgcaaaacca ccattgggga cagggaggtg
     1561 gattctgatg cctactatgt ctacagactc caggtgtcat ccatcaacgt ctctgtgaac
     1621 gcagtgcaga ctgtggtccg ccagggtgag aacatcaccc tcatgtgcat tgtgatcggg
     1681 aatgaggtgg tcaacttcga gtggacatac ccccgcaaag aaagtgggcg gctggtggag
     1741 ccggtgactg acttcctctt ggatatgcct taccacatcc gctccatcct gcacatcccc
     1801 agtgccgagt tagaagactc ggggacctac acctgcaatg tgacggagag tgtgaatgac
     1861 catcaggatg aaaaggccat caacatcacc gtggttgaga gcggctacgt gcggctcctg
     1921 ggagaggtgg gcacactaca atttgctgag ctgcatcgga gccggacact gcaggtagtg
     1981 ttcgaggcct acccaccgcc cactgtcctg tggttcaaag acaaccgcac cctgggcgac
     2041 tccagcgctg gcgaaatcgc cctgtccacg cgcaacgtgt cggagacccg gtatgtgtca
     2101 gagctgacac tggttcgcgt gaaggtggca gaggctggcc actacaccat gcgggccttc
     2161 catgaggatg ctgaggtcca gctctccttc cagctacaga tcaatgtccc tgtccgagtg
     2221 ctggagctaa gtgagagcca ccctgacagt ggggaacaga cagtccgctg tcgtggccgg
     2281 ggcatgcccc agccgaacat catctggtct gcctgcagag acctcaaaag gtgtccacgt
     2341 gagctgccgc ccacgctgct ggggaacagt tccgaagagg agagccagct ggagactaac
     2401 gtgacgtact gggaggagga gcaggagttt gaggtggtga gcacactgcg tctgcagcac
     2461 gtggatcggc cactgtcggt gcgctgcacg ctgcgcaacg ctgtgggcca ggacacgcag
     2521 gaggtcatcg tggtgccaca ctccttgccc tttaaggtgg tggtgatctc agccatcctg
     2581 gccctggtgg tgctcaccat catctccctt atcatcctca tcatgctttg gcagaagaag
     2641 ccacgttacg agatccgatg gaaggtgatt gagtctgtga gctctgacgg ccatgagtac
     2701 atctacgtgg accccatgca gctgccctat gactccacgt gggagctgcc gcgggaccag
     2761 cttgtgctgg gacgcaccct cggctctggg gcctttgggc aggtggtgga ggccacggct
     2821 catggcctga gccattctca ggccacgatg aaagtggccg tcaagatgct taaatccaca
     2881 gcccgcagca gtgagaagca agcccttatg tcggagctga agatcatgag tcaccttggg
     2941 ccccacctga acgtggtcaa cctgttgggg gcctgcacca aaggaggacc catctatatc
     3001 atcactgagt actgccgcta cggagacctg gtggactacc tgcaccgcaa caaacacacc
     3061 ttcctgcagc accactccga caagcgccgc ccgcccagcg cggagctcta cagcaatgct
     3121 ctgcccgttg ggctccccct gcccagccat gtgtccttga ccggggagag cgacggtggc
     3181 tacatggaca tgagcaagga cgagtcggtg gactatgtgc ccatgctgga catgaaagga
     3241 gacgtcaaat atgcagacat cgagtcctcc aactacatgg ccccttacga taactacgtt
     3301 ccctctgccc ctgagaggac ctgccgagca actttgatca acgagtctcc agtgctaagc
     3361 tacatggacc tcgtgggctt cagctaccag gtggccaatg gcatggagtt tctggcctcc
     3421 aagaactgcg tccacagaga cctggcggct aggaacgtgc tcatctgtga aggcaagctg
     3481 gtcaagatct gtgactttgg cctggctcga gacatcatgc gggactcgaa ttacatctcc
     3541 aaaggcagca cctttttgcc tttaaagtgg atggctccgg agagcatctt caacagcctc
     3601 tacaccaccc tgagcgacgt gtggtccttc gggatcctgc tctgggagat cttcaccttg
     3661 ggtggcaccc cttacccaga gctgcccatg aacgagcagt tctacaatgc catcaaacgg
     3721 ggttaccgca tggcccagcc tgcccatgcc tccgacgaga tctatgagat catgcagaag
     3781 tgctgggaag agaagtttga gattcggccc cccttctccc agctggtgct gcttctcgag
     3841 agactgttgg gcgaaggtta caaaaagaag taccagcagg tggatgagga gtttctgagg
     3901 agtgaccacc cagccatcct tcggtcccag gcccgcttgc ctgggttcca tggcctccga
     3961 tctcccctgg acaccagctc cgtcctctat actgccgtgc agcccaatga gggtgacaac
     4021 gactatatca tccccctgcc tgaccccaaa cccgaggttg ctgacgaggg cccactggag
     4081 ggttccccca gcctagccag ctccaccctg aatgaagtca acacctcctc aaccatctcc
     4141 tgtgacagcc ccctggagcc ccaggacgaa ccagagccag agccccagct tgagctccag
     4201 gtggagccgg agccagagct ggaacagttg ccggattcgg ggtgccctgc gcctcgggcg
     4261 gaagcagagg atagcttcct gctcgaggac tacaaagacc atgacggtga ttataaagat
     4321 catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta aacccgctga
     4381 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4441 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4501 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4561 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4621 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4681 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4741 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4801 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4861 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4921 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4981 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5041 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5101 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5161 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5221 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5281 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5341 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5401 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5461 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5521 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5581 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5641 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5701 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5761 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5821 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5881 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5941 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6001 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6061 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6121 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6181 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6241 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6301 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6361 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6421 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6481 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6541 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6601 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6661 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6721 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6781 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6841 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6901 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6961 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7021 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7081 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7141 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7201 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7261 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7321 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7381 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7441 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7501 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7561 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7621 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7681 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7741 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7801 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7861 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7921 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7981 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8041 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8101 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8161 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8221 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8281 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8341 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8401 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8461 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8521 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8581 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8641 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8701 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8761 cacatttccc cgaaaagtgc cacctgacgt c
//
