LOCUS       V042499                 8338 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042499
VERSION     V042499
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..3828
                     /label="MIB2(NM_080875)"
                     /note="MIB2(NM_080875)"
                     /gene="MIB2"
     CDS             3835..3900
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3938..4162
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4208..4636
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4650..4979
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5046..5840
                     /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    6014..6135
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6184..6200)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6208..6224
                     /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(6232..6262)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6277..6298
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6586..7171)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7342..8202)
                     /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(8203..8307)
                     /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 accatggacc cagaccccca ggcgggcgtg caggtgggca tgcgggtggt gcgcggcgtg
     1021 gactggaagt ggggccagca ggacggcggc gagggcggcg tgggcacggt ggtggagctt
     1081 ggccgccacg gcagcccctc gacacccgac cgcacagtgg tcgtgcagtg ggaccagggc
     1141 acgcgcacca actaccgcgc cggctaccag ggcgcgcacg acctgctgct gtacgacaac
     1201 gcccagatcg gcgtccggca ccccaacatc atctgtgact gctgcaagaa gcacgggctg
     1261 cgggggatgc gctggaagtg ccgtgtgtgc ctggactacg acctctgcac gcagtgctac
     1321 atgcacaaca agcatgagct cgcccacgcc ttcgaccgct acgagaccgc tcactcgcgc
     1381 cctgtcacac tgagtccccg ccagggcctc ccgaggatcc cactaagggg catcttccag
     1441 ggagcgaagg tggtgcgagg ccccgactgg gagtggggct cacaggatgg aggggaaggg
     1501 aaaccgggcc gtgtggtgga catccgtggc tgggatgtgg agacaggccg gagtgtggcc
     1561 agcgtgacgt gggctgatgg taccaccaat gtgtaccgtg tgggccacaa gggcaaggtg
     1621 gacctcaagt gtgtgggcga ggcagcgggc ggcttctact acaaggacca cctcccaagg
     1681 ctcggcaagc cggcggagct gcagcgcagg gtgagtgctg acagccagcc cttccagcac
     1741 ggggacaagg tcaagtgtct gctggacact gatgtcctgc gggagatgca ggaaggccac
     1801 ggcggctgga accccaggat ggcggagttt atcggacaga cgggcaccgt gcatcgtatc
     1861 acggaccgcg gggacgtgcg cgtgcagttc aaccacgaga cgcgctggac cttccacccc
     1921 ggggcgctca ccaagcacca ctccttctgg gtgggcgacg tggtccgggt catcggcgac
     1981 cttgacacag tgaagcggct gcaggctggg catggcgagt ggacggacga catggcccct
     2041 gccctgggcc gcgtcgggaa ggtggtgaaa gtgtttggag acgggaacct gcgtgtagca
     2101 gtcgctggtc agcggtggac cttcagcccc tcctgcctgg tggcctaccg gcccgaggag
     2161 gatgccaacc tggacgtggc cgagcgcgcc cgggagaaca aaagctcact gagcgtggcc
     2221 ctggacaagc ttcgggccca gaagagtgac ccagagcacc cgggaaggct ggtggtggag
     2281 gtggcgctgg gtaacgcagc ccgggctctg gacctgctgc ggaggcgccc agagcaggtg
     2341 gacaccaaga accaaggcag gaccgctctg caagtggctg cctacctggg ccaggtggag
     2401 ttgatacggc tgctgctaca agccagggcg ggcgtggacc tgccggacga cgagggcaac
     2461 acggcactgc actacgcggc cctggggaac cagcccgagg ccaccagggt gctcctgagt
     2521 gctgggtgcc gggcggacgc catcaacagc acccagagca cagcactgca cgtggccgtg
     2581 cagaggggct tcctggaggt ggtgcgggcc ctgtgtgagc gcggctgtga cgtcaacctg
     2641 cccgacgccc actcggacac gcccctgcac tccgccatct cggcgggcac tggagccagc
     2701 ggcattgtcg aggtcctcac ggaggtgcca aacatcgatg ttaccgccac caacagccag
     2761 ggtttcaccc tgctgcacca tgcctccctc aagggtcacg cgctagctgt gagaaagatt
     2821 ctggctcggg cgcggcagct ggtggacgcc aagaaggagg acggcttcac ggcgctgcat
     2881 ctggctgccc tcaacaacca ccgcgaggtg gcccagatcc tcatccggga gggccgctgt
     2941 gacgtgaacg tgcgcaaccg gaagctgcag tccccgctgc atctcgccgt gcaacaggcc
     3001 cacgtggggc tggtgccgct actggtggac gctgggtgca gtgtcaacgc cgaggacgag
     3061 gagggggaca cagccctgca cgtggcgctg cagcgtcatc agctgctgcc cctggtggct
     3121 gatggggccg ggggggaccc agggcccttg cagctgctgt ccaggctaca ggcctcgggc
     3181 ctccccggca gcgcggagct gacggtgggc gcggcggtcg cctgcttcct ggcgctggag
     3241 ggcgccgacg tgagctacac caaccaccgc ggtcggagcc cgctggacct ggccgccgag
     3301 ggtcgcgtgc tcaaggccct tcagggctgc gcccagcgct tccgggagcg gcaggcgggc
     3361 gggggcgcgg ccccgggccc caggcaaacg ctcgggaccc ccaacaccgt gacgaacctg
     3421 cacgtgggcg ccgcgccggg gcccgaggcc gctgagtgcc tggtgtgctc cgagctggcg
     3481 ctgctggtgc tgttctcgcc gtgccagcac cgcaccgtgt gtgaggagtg cgcgcgcagg
     3541 atgaagaagt gcatcaggtg ccaggtggtc gtcagcaaga aactgcgccc agacggctct
     3601 gaggtggcga gcgccgcccc cgcccccggc ccgccgcgcc agctggtgga ggagctgcag
     3661 agccgctacc ggcagatgga ggaacgcatc acctgcccca tctgcatcga cagccacatc
     3721 cgcctcgtgt tccagtgcgg ccacggcgca tgcgccccct gcggctccgc gctcagcgcc
     3781 tgccccatct gccgccagcc catccgcgac cgcatccaga tcttcgtgct cgaggactac
     3841 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     3901 tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3961 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4021 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4081 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4141 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4201 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4261 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4321 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4381 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4441 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4561 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4621 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4681 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4741 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4801 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4861 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4921 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4981 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5041 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5101 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5161 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5221 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5281 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5341 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5401 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5461 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5521 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5581 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5641 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5701 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5761 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5821 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5881 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5941 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6001 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6061 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6121 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6181 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6361 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6421 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6481 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6541 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     7141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7321 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7381 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7441 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7501 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7561 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7621 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7681 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7741 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7801 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7861 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7921 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7981 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8041 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8101 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8161 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8221 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8281 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
