LOCUS       V020792                 9341 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020792
VERSION     V020792
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..4831
                     /label="INSRR(NM_014215)"
                     /note="INSRR(NM_014215)"
                     /gene="INSRR"
     CDS             4838..4903
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4941..5165
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5211..5639
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5653..5982
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6049..6843
                     /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    7017..7138
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7187..7203)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7211..7227
                     /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(7235..7265)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7280..7301
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7589..8174)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8345..9205)
                     /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(9206..9310)
                     /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 atggcagtgc ctagtctgtg
      961 gccctgggga gcatgcctgc ctgtgatctt cctctccttg ggatttggcc tggatacagt
     1021 agaggtgtgc cccagcctgg atattcgctc agaggtggca gagcttcgtc agctggagaa
     1081 ctgcagcgtg gtggagggcc acctgcagat cctgctcatg ttcacagcca ccggggagga
     1141 cttccgcggc ctcagcttcc ctcgcctcac ccaggtcacc gactacctgc tgctcttccg
     1201 tgtctacgga ctggagagcc tgcgcgacct cttccccaac ctagcagtca tccgcgggac
     1261 gcgcctcttc ctgggctatg cactggtcat ctttgagatg ccacatctgc gtgacgtggc
     1321 actgcctgca cttggggccg tgctgcgtgg ggctgtgcgt gtggagaaga accaggagct
     1381 ctgccacctc tccaccattg actggggact gctgcagcca gcacctggcg ccaaccacat
     1441 cgtgggcaac aagctgggcg aggagtgtgc tgacgtgtgc cctggtgtgc tgggtgctgc
     1501 tggtgagccc tgtgccaaga ccaccttcag cgggcacact gactacagat gctggacctc
     1561 cagccactgc cagagagtgt gcccctgccc ccatgggatg gcttgcacag cgaggggcga
     1621 gtgctgccac accgaatgcc tggggggctg cagccagcca gaagaccctc gtgcctgtgt
     1681 agcttgccgc cacctctact tccagggtgc ctgcctgtgg gcctgcccgc caggcaccta
     1741 ccagtatgag tcctggcgct gtgtcacagc tgagcgctgt gccagcctgc actctgtgcc
     1801 cggccgtgcc tccaccttcg gcatacacca gggcagttgc ctggcccagt gcccttctgg
     1861 cttcacccgt aatagcagca gcatattctg ccacaagtgc gaggggctgt gccctaaaga
     1921 gtgcaaggta ggcaccaaga ccatcgactc catccaggcg gcacaggatc ttgtgggctg
     1981 cacgcatgtg gagggaagcc tcatcctcaa ccttcgccag ggctacaacc tggagccaca
     2041 gctgcagcac agcctggggc tggtagaaac cattactggc ttcctcaaaa tcaagcactc
     2101 ctttgccctc gtgtccctgg gctttttcaa gaacctcaaa ctaatccggg gagacgccat
     2161 ggtggatggg aactacactc tctacgtgct ggacaaccag aacctacaac agctagggtc
     2221 ctgggtggcc gcggggctca ccattcccgt gggcaagatc tacttcgcct tcaacccgcg
     2281 cctctgcttg gaacacatct accgactgga ggaggtgaca ggcacgcgag gtcggcagaa
     2341 caaggctgag atcaaccccc gcaccaacgg agaccgcgcc gcctgccaga ctcgcaccct
     2401 gcgcttcgtg tccaacgtga cggaggcaga ccgcatcctg ctacgctggg agcgctatga
     2461 gccactggag gcccgcgacc tgctcagctt catcgtgtac tacaaggagt ccccattcca
     2521 gaacgccaca gagcacgtgg gtccagatgc ttgtggaacc cagagctgga acctgctgga
     2581 tgtggagctg cccctaagcc gcacccagga gccaggggtg accctagcct ccctcaagcc
     2641 ttggacacag tacgcagtgt ttgtgcgggc catcacgcta accactgagg aggacagccc
     2701 tcatcaagga gcccagagtc ccatcgtcta cctccgaacg ctgcctgcag ctcccacggt
     2761 gccccaagac gtcatctcca cgtccaactc ctcctcccac ctcctggtgc gctggaagcc
     2821 accgacccag cgcaatggga acctcaccta ctacctggtg ctgtggcagc ggctggcaga
     2881 ggacggcgac ctctacctca atgactactg ccaccgcggc ttgcggctgc ccaccagcaa
     2941 caacgatccg cgcttcgacg gcgaagacgg ggatcctgag gccgagatgg agtccgactg
     3001 ctgcccttgc cagcacccac ctcctggtca ggttctgccc ccgctggagg cgcaagaggc
     3061 ctcgttccag aagaagtttg aaaactttct acacaacgcg atcaccatcc ccatatcccc
     3121 ttggaaggtg acgtccatca acaagagccc ccaaagggac tcagggcggc accgccgggc
     3181 agctgggccc ctccggctgg ggggcaacag ctcggatttc gagatccagg aggacaaggt
     3241 gccccgtgag cgagcggtgc tgagcggcct gcgccacttc acggaatacc ggatcgacat
     3301 ccatgcctgc aaccacgcgg cgcacaccgt gggctgcagc gccgccacct tcgtctttgc
     3361 gcgcaccatg ccccacagag aggctgatgg tattccagga aaggtggcct gggaggcctc
     3421 cagcaagaac agtgtccttc tgcgctggct cgagccacca gaccccaacg gactcatcct
     3481 caagtacgaa atcaagtacc gccgcttggg agaggaggcc acagtgctgt gtgtgtcccg
     3541 tcttcgatat gcgaagtttg ggggagtcca cctggccctg ctgccccctg gaaactactc
     3601 tgccagggtt agggcaacct cactggctgg caatggctct tggacagaca gtgttgcctt
     3661 ctacatcctt ggcccagagg aggaggatgc tggggggctg catgtcctcc tcactgccac
     3721 ccctgtgggg ctcacgctgc tcatcgttct tgctgccctt ggtttcttct acggcaagaa
     3781 gagaaacaga accctgtatg cttctgtgaa tccagagtac ttcagcgcct ctgatatgta
     3841 tgtccctgat gaatgggagg tgcctcggga gcagatctcg ataatccggg aactgggcca
     3901 gggctctttt gggatggtat atgaggggct ggcacgagga cttgaggctg gagaggagtc
     3961 cacacccgtg gccctgaaga cggtgaatga gctggccagc ccacgggaat gcattgagtt
     4021 cctcaaggaa gcttctgtca tgaaagcctt caagtgtcac catgtggtgc gtctcctggg
     4081 tgtggtatct cagggccagc caactctggt catcatggag ttaatgaccc gtggggacct
     4141 caagagccat cttcgatctt tgcggcctga ggcagagaac aaccctgggc tcccacagcc
     4201 agcattgggg gaaatgatcc aaatggctgg tgagattgca gacggcatgg cctaccttgc
     4261 tgccaacaag tttgtgcacc gagatctagc agcccgcaac tgcatggtgt cccaggactt
     4321 caccgtcaag atcggggact tcgggatgac tcgggacgtg tatgagacag actattaccg
     4381 caagggtggg aaggggctgc tgcccgtgcg ctggatggcc cccgagtccc tcaaagatgg
     4441 gatcttcacc acccactcgg atgtctggtc ctttggcgtg gtactctggg agattgtgac
     4501 cctggcagaa caaccctacc agggcctgtc caatgagcag gtgctgaagt tcgtcatgga
     4561 tggcggggtc ctggaggagc tggagggctg tccccttcag ctgcaggagc tgatgagccg
     4621 ctgctggcag ccgaacccac gcctgcgccc atctttcaca cacattctgg acagcataca
     4681 ggaggagctg cggccctcct tccgcctcct ctccttctac tacagcccgg aatgccgggg
     4741 ggcccggggc tccctgccta ccaccgatgc agagcctgac tcctcaccca ctccaagaga
     4801 ctgcagccct caaaatgggg gtccagggca cctcgaggac tacaaagacc atgacggtga
     4861 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     4921 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4981 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     5041 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     5101 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     5161 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     5221 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5281 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5341 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5401 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5461 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5521 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5581 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     5641 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5701 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5821 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5881 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5941 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     6001 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     6061 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6121 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6181 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6241 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6301 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6361 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6421 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6481 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6541 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6601 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6661 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6721 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6781 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6841 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6901 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6961 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     7021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7081 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7141 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     7201 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7261 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7321 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7381 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7441 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7501 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7561 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7621 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7681 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7741 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7801 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7861 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7921 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7981 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     8041 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     8101 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8161 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8221 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8281 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8341 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8401 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8461 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8521 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8581 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8641 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8701 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8761 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8821 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8881 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8941 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9001 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9061 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9181 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9241 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9301 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
