LOCUS       V020337                 9551 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020337
VERSION     V020337
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             941..5041
                     /label="IGF1R(NM_000875)"
                     /note="IGF1R(NM_000875)"
                     /gene="IGF1R"
     CDS             5048..5113
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5151..5375
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5421..5849
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5863..6192
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6259..7053
                     /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    7227..7348
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7397..7413)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7421..7437
                     /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(7445..7475)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7490..7511
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7799..8384)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8555..9415)
                     /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(9416..9520)
                     /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 atccgccacc atgaagtctg gctccggagg
      961 agggtccccg acctcgctgt gggggctcct gtttctctcc gccgcgctct cgctctggcc
     1021 gacgagtgga gaaatctgcg ggccaggcat cgacatccgc aacgactatc agcagctgaa
     1081 gcgcctggag aactgcacgg tgatcgaggg ctacctccac atcctgctca tctccaaggc
     1141 cgaggactac cgcagctacc gcttccccaa gctcacggtc attaccgagt acttgctgct
     1201 gttccgagtg gctggcctcg agagcctcgg agacctcttc cccaacctca cggtcatccg
     1261 cggctggaaa ctcttctaca actacgccct ggtcatcttc gagatgacca atctcaagga
     1321 tattgggctt tacaacctga ggaacattac tcggggggcc atcaggattg agaaaaatgc
     1381 tgacctctgt tacctctcca ctgtggactg gtccctgatc ctggatgcgg tgtccaataa
     1441 ctacattgtg gggaataagc ccccaaagga atgtggggac ctgtgtccag ggaccatgga
     1501 ggagaagccg atgtgtgaga agaccaccat caacaatgag tacaactacc gctgctggac
     1561 cacaaaccgc tgccagaaaa tgtgcccaag cacgtgtggg aagcgggcgt gcaccgagaa
     1621 caatgagtgc tgccaccccg agtgcctggg cagctgcagc gcgcctgaca acgacacggc
     1681 ctgtgtagct tgccgccact actactatgc cggtgtctgt gtgcctgcct gcccgcccaa
     1741 cacctacagg tttgagggct ggcgctgtgt ggaccgtgac ttctgcgcca acatcctcag
     1801 cgccgagagc agcgactccg aggggtttgt gatccacgac ggcgagtgca tgcaggagtg
     1861 cccctcgggc ttcatccgca acggcagcca gagcatgtac tgcatccctt gtgaaggtcc
     1921 ttgcccgaag gtctgtgagg aagaaaagaa aacaaagacc attgattctg ttacttctgc
     1981 tcagatgctc caaggatgca ccatcttcaa gggcaatttg ctcattaaca tccgacgggg
     2041 gaataacatt gcttcagagc tggagaactt catggggctc atcgaggtgg tgacgggcta
     2101 cgtgaagatc cgccattctc atgccttggt ctccttgtcc ttcctaaaaa accttcgcct
     2161 catcctagga gaggagcagc tagaagggaa ttactccttc tacgtcctcg acaaccagaa
     2221 cttgcagcaa ctgtgggact gggaccaccg caacctgacc atcaaagcag ggaaaatgta
     2281 ctttgctttc aatcccaaat tatgtgtttc cgaaatttac cgcatggagg aagtgacggg
     2341 gactaaaggg cgccaaagca aaggggacat aaacaccagg aacaacgggg agagagcctc
     2401 ctgtgaaagt gacgtcctgc atttcacctc caccaccacg tcgaagaatc gcatcatcat
     2461 aacctggcac cggtaccggc cccctgacta cagggatctc atcagcttca ccgtttacta
     2521 caaggaagca ccctttaaga atgtcacaga gtatgatggg caggatgcct gcggctccaa
     2581 cagctggaac atggtggacg tggacctccc gcccaacaag gacgtggagc ccggcatctt
     2641 actacatggg ctgaagccct ggactcagta cgccgtttac gtcaaggctg tgaccctcac
     2701 catggtggag aacgaccata tccgtggggc caagagtgag atcttgtaca ttcgcaccaa
     2761 tgcttcagtt ccttccattc ccttggacgt tctttcagca tcgaactcct cttctcagtt
     2821 aatcgtgaag tggaaccctc cctctctgcc caacggcaac ctgagttact acattgtgcg
     2881 ctggcagcgg cagcctcagg acggctacct ttaccggcac aattactgct ccaaagacaa
     2941 aatccccatc aggaagtatg ccgacggcac catcgacatt gaggaggtca cagagaaccc
     3001 caagactgag gtgtgtggtg gggagaaagg gccttgctgc gcctgcccca aaactgaagc
     3061 cgagaagcag gccgagaagg aggaggctga ataccgcaaa gtctttgaga atttcctgca
     3121 caactccatc ttcgtgccca gacctgaaag gaagcggaga gatgtcatgc aagtggccaa
     3181 caccaccatg tccagccgaa gcaggaacac cacggccgca gacacctaca acatcaccga
     3241 cccggaagag ctggagacag agtacccttt ctttgagagc agagtggata acaaggagag
     3301 aactgtcatt tctaaccttc ggcctttcac attgtaccgc atcgatatcc acagctgcaa
     3361 ccacgaggct gagaagctgg gctgcagcgc ctccaacttc gtctttgcaa ggactatgcc
     3421 cgcagaagga gcagatgaca ttcctgggcc agtgacctgg gagccaaggc ctgaaaactc
     3481 catcttttta aagtggccgg aacctgagaa tcccaatgga ttgattctaa tgtatgaaat
     3541 aaaatacgga tcacaagttg aggatcagcg agaatgtgtg tccagacagg aatacaggaa
     3601 gtatggaggg gccaagctaa accggctaaa cccggggaac tacacagccc ggattcaggc
     3661 cacatctctc tctgggaatg ggtcgtggac agatcctgtg ttcttctatg tccaggccaa
     3721 aacaggatat gaaaacttca tccatctgat catcgctctg cccgtcgctg tcctgttgat
     3781 cgtgggaggg ttggtgatta tgctgtacgt cttccataga aagagaaata acagcaggct
     3841 ggggaatgga gtgctgtatg cctctgtgaa cccggagtac ttcagcgctg ctgatgtgta
     3901 cgttcctgat gagtgggagg tggctcggga gaagatcacc atgagccggg aacttgggca
     3961 ggggtcgttt gggatggtct atgaaggagt tgccaagggt gtggtgaaag atgaacctga
     4021 aaccagagtg gccattaaaa cagtgaacga ggccgcaagc atgcgtgaga ggattgagtt
     4081 tctcaacgaa gcttctgtga tgaaggagtt caattgtcac catgtggtgc gattgctggg
     4141 tgtggtgtcc caaggccagc caacactggt catcatggaa ctgatgacac ggggcgatct
     4201 caaaagttat ctccggtctc tgaggccaga aatggagaat aatccagtcc tagcacctcc
     4261 aagcctgagc aagatgattc agatggccgg agagattgca gacggcatgg catacctcaa
     4321 cgccaataag ttcgtccaca gagaccttgc tgcccggaat tgcatggtag ccgaagattt
     4381 cacagtcaaa atcggagatt ttggtatgac gcgagatatc tatgagacag actattaccg
     4441 gaaaggaggg aaagggctgc tgcccgtgcg ctggatgtct cctgagtccc tcaaggatgg
     4501 agtcttcacc acttactcgg acgtctggtc cttcggggtc gtcctctggg agatcgccac
     4561 actggccgag cagccctacc agggcttgtc caacgagcaa gtccttcgct tcgtcatgga
     4621 gggcggcctt ctggacaagc cagacaactg tcctgacatg ctgtttgaac tgatgcgcat
     4681 gtgctggcag tataacccca agatgaggcc ttccttcctg gagatcatca gcagcatcaa
     4741 agaggagatg gagcctggct tccgggaggt ctccttctac tacagcgagg agaacaagct
     4801 gcccgagccg gaggagctgg acctggagcc agagaacatg gagagcgtcc ccctggaccc
     4861 ctcggcctcc tcgtcctccc tgccactgcc cgacagacac tcaggacaca aggccgagaa
     4921 cggccccggc cctggggtgc tggtcctccg cgccagcttc gacgagagac agccttacgc
     4981 ccacatgaac gggggccgca agaacgagcg ggccttgccg ctgccccagt cttcgacctg
     5041 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     5101 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     5161 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5221 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5281 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5341 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5401 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5461 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5521 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5581 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5641 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5701 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5761 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5821 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5941 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6001 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6061 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6121 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6181 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     6241 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6301 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6361 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6421 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6481 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6541 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6601 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6661 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6721 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6781 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6841 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6901 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6961 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7021 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7081 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     7141 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     7201 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     7261 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7321 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     7381 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7441 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7501 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7561 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7621 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7681 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7741 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     8341 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8401 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8461 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8521 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8581 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8641 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8701 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8761 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8821 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8881 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8941 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9001 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9061 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9121 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9181 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9241 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9301 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9361 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9421 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9481 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9541 cacctgacgt c
//
