LOCUS       V042033                 9369 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042033
VERSION     V042033
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4049
                     /label="Pdgfrb(NM_001146268)"
                     /note="Pdgfrb(NM_001146268)"
                     /gene="Pdgfrb"
     CDS             4056..4121
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4145..4733
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4799..4847
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5049..5477
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5491..5820
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5887..6681
                     /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    6857..6978
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7027..7043)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7051..7067
                     /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(7075..7105)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7120..7141
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7429..8017)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8188..9048)
                     /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(9049..9153)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg gcttccagga gtgataccag
      781 ctttagtcct cagaggccag ttgttgctgt ccgtgttatg gctcctggga ccgcagacct
      841 cccggggcct agtcatcacg ccccctgggc cagagtttgt tctcaacatc tccagcacct
      901 ttgttctgac ctgctcgggc tcagctccgg tgatgtggga acagatgtcc caggtgccct
      961 ggcaagaagc ggccatgaat caggacggca ccttctccag tgtgctgaca ctgaccaatg
     1021 tcactggggg agacactggg gaatactttt gtgtctacaa caactcacta gggccggagc
     1081 tcagtgagag gaagcgtatc tatatctttg tgccagatcc caccatgggc ttcctcccta
     1141 tggactccga ggacctgttc atttttgtca cggatgtcac tgagacgaca attccgtgcc
     1201 gagtgacaga cccccagctg gaggtgacgc tacatgagaa gaaagtggat atccccctac
     1261 acgtacccta cgaccaccag cgaggtttca ctggtacttt tgaggacaag acttacattt
     1321 gcaaaaccac cattggggac agggaagtgg actccgatac ttactacgtc tacagcctcc
     1381 aggtgtcatc catcaacgtc tctgtgaatg ccgtgcagac tgtggtccgc cagggcgaga
     1441 gcatcaccat ccggtgcatt gtgatgggca atgatgtggt gaacttccaa tggacgtacc
     1501 cccgcatgaa gagtgggcgg ctggtggagc cagtgacaga ctacctcttt ggagtgccct
     1561 cccgcattgg ctccatcctg catatcccca cggctgagct gagtgattcg ggcacctata
     1621 cttgcaacgt gtcagtgagt gtgaacgacc atggcgatga gaaagccatc aacatctctg
     1681 tgatcgagaa tggctacgtg cggctgctgg agacactggg agatgtagaa attgctgagc
     1741 tgcaccggag tcggacgctg cgggtggtgt tcgaggctta tccgatgcct tctgtcctgt
     1801 ggctcaagga caaccgtacc ttgggtgact ccggcgctgg cgagttagtt ttgtctactc
     1861 gcaacatgtc tgagacccgg tacgtgtcag aactgatcct ggtacgtgtg aaggtgtcag
     1921 aagcgggcta ctatactatg cgagccttcc acgaggacga tgaggtccag ctctccttca
     1981 agctgcaggt caatgtcccc gtccgtgtgc tggagctgag tgagagtcac cctgccaatg
     2041 gggagcagac aatccgctgt cgtggccggg gcatgcctca gccaaatgtc acctggtcta
     2101 cctgcagaga cctcaaaagg tgtccacgaa aactgtcacc cacacccttg gggaatagtt
     2161 ccaaggagga gagccagcta gaaacgaatg tgactttctg ggaggaagat caggaatacg
     2221 aggtggtgag cacactgcgc ctgcgccacg tggatcagcc actgtccgta cgctgcatgc
     2281 tgcagaactc catgggtgga gattcgcagg aggtcaccgt ggtcccacat tccttgccct
     2341 tcaaagtggt ggtgatctca gccatcctgg ccttagtggt ccttaccgtc atctctctca
     2401 tcatcctcat catgctgtgg cagaagaagc cacgctatga gatccgatgg aaggtcattg
     2461 agtctgtgag ctctgacggt catgagtaca tctacgtgga ccctgtgcag ttgccttacg
     2521 actccacctg ggagctgcca cgggaccagc ttgttctggg acgcactctt ggctctgggg
     2581 ctttcggaca ggtggtggag gccacagctc acggtctgag ccattcgcag gccaccatga
     2641 aagtggctgt caagatgctg aaatcgacag ccagaagtag cgagaagcaa gccttaatgt
     2701 ccgagctgaa gattatgagt catcttggac cccacctgaa cgtggtcaac ctgctggggg
     2761 cctgcaccaa aggagggccc atctacatca tcacggaata ctgccgatac ggtgatctgg
     2821 tggactacct gcaccggaac aaacacacct tcttgcagcg acactccaac aagcattgtc
     2881 cgcccagtgc tgagctctac agcaacgccc tgccagtggg gttctcccta cccagccact
     2941 tgaacctgac tggggagagt gacggtggct acatggatat gagcaaggat gaatctatag
     3001 attacgtgcc catgttggac atgaaaggag acatcaaata cgcagacatt gagtccccca
     3061 gctacatggc cccttatgat aactatgtcc catctgcccc tgaaaggacc tatcgcgcca
     3121 ccttaatcaa cgactcacca gtgctcagct acacagacct cgtgggcttc agctaccaag
     3181 tggccaacgg catggacttc ttagcctcta agaactgtgt tcaccgagac ttggcggcca
     3241 ggaatgtgct catctgcgag ggcaagctgg tcaagatctg tgacttcggc ctggctcgag
     3301 acatcatgag ggactcaaac tacatctcca aaggcagcac cttcctgcct ctgaagtgga
     3361 tggccccaga gagcatcttc aacagcctct acaccacttt gagtgatgtc tggtcttttg
     3421 ggatcctact ctgggagatc ttcacactgg gtggcacccc ttacccagag ctgcccatga
     3481 acgaccagtt ctacaatgcc atcaagaggg gctaccgcat ggcccagcct gctcatgcct
     3541 ccgacgagat ctatgagatc atgcagaaat gctgggaaga aaagtttgag actcgacccc
     3601 ccttctccca gctggtgctg ctcctggaga ggcttctggg tgaaggctat aaaaagaagt
     3661 accagcaggt agatgaggag ttcctgagga gtgaccatcc tgccatcctg aggtcccaag
     3721 cccgctttcc ggggatccac agcctccgat cccctctgga caccagctct gttctctaca
     3781 ctgccgtgca gcccaatgag agtgacaatg actacatcat ccccttacct gaccccaagc
     3841 ctgacgttgc tgatgaaggt ctcccagagg ggtcccccag ccttgccagt tccaccttga
     3901 atgaagtcaa cacttcctcc accatctcct gcgacagtcc cctggagctc caagaagagc
     3961 cacagcaagc agagcctgag gcacaactgg agcagccaca ggattcaggc tgcccaggac
     4021 ctctggctga agcagaggac agcttcctgg gatccgacta caaagaccat gacggtgatt
     4081 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     4141 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4201 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4261 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4321 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4381 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4441 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4501 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4561 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4621 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4681 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     4741 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4801 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     4861 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4921 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4981 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     5041 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5101 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5161 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     5221 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5281 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5341 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5401 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     5461 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5521 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5581 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5641 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5701 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5761 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5821 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5881 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5941 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6001 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6061 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6121 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6181 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6241 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6301 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6361 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6421 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6481 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6541 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6601 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6661 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     6721 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6781 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6841 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6901 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6961 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7021 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7081 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7141 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7201 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7261 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7321 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7381 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7441 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7501 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7561 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7621 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7681 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7741 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7801 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7861 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7921 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8101 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8161 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8221 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8281 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8341 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8401 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8461 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8521 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8581 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8641 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8701 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8761 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8821 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8881 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8941 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9001 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9061 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9181 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     9241 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     9301 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     9361 ctgcttagg
//
