BBa_I713001 1 BBa_I713001 A pBBR1MCS plasmid without the mob gene. 2007-10-24T11:00:00Z 2015-08-31T04:07:47Z Plasmid: pBBR1MCS Primer: DMOBF: AATTGCTAGCTGACGTACTCGACCGCCAACACAGC DMOBR: AATAGCTAGCATCCCAAGAGACGGCCCGAGACCTTC LOCUS pBBR1MCS 4707 bp DNA circular SYN 4-JUN-2007 DEFINITION Cloning vector pBBR1MCS, complete sequence. ACCESSION U02374 SOURCE Cloning vector pBBR1MCS. ORGANISM Cloning vector pBBR1MCS artificial sequence; vectors. REFERENCE 1 (bases 1 to 4707) AUTHORS Kovach,M.E., Phillips,R.W., Elzer,P.H., Roop,R.M. 2nd and Peterson,K.M. TITLE pBBR1MCS: a broad-host-range cloning vector JOURNAL BioTechniques 16 (5), 800-802 (1994) MEDLINE 94347360 REFERENCE 2 (bases 1 to 4707) AUTHORS Kovach,M.E., Elzer,P.H., Hill,D.S., Robertson,G.T., Farris,M.A., Roop,R.M. II and Peterson,K.M. TITLE Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes JOURNAL Gene 166 (1), 175-176 (1995) MEDLINE 96105222 REFERENCE 3 (bases 1 to 4707) AUTHORS Kovach,M.E. TITLE Direct Submission JOURNAL Submitted (05-OCT-1993) Michael E. Kovach, Microbiology & Immunology, Louisiana State University Medical Center-Shreveport, 1501 Kings Highway, Shreveport, LA 71130, USA BASE COUNT 966 a 1380 c 1329 g 1032 t ORIGIN 1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2581 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2641 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2701 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2761 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2821 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2881 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2941 agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3001 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3181 gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc 3241 tagaactagt ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga 3301 cctcgagggg gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct 3361 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 3421 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 3481 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 3541 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgcatgcata 3601 aaaactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat 3661 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg 3721 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac 3781 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg 3841 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat 3901 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt 3961 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt 4021 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct 4081 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg 4141 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat 4201 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc 4261 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac 4321 ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat 4381 caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt 4441 cgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg 4501 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcgcccgcgt tcctgctggc 4561 gctgggcctg tttctggcgc tggacttccc gctgttccgt cagcagcttt tcgcccacgg 4621 ccttgatgat cgcggcggcc ttggcctgca tatcccgatt caacggcccc agggcgtcca 4681 gaacgggctt caggcgctcc cgaaggt // This plasmid is constructed by PCR with the pBBR1MCS plasmid as template and the following primers: DMOBF: AATTGCTAGCTGACGTACTCGACCGCCAACACAGC DMOBR: AATAGCTAGCATCCCAAGAGACGGCCCGAGACCTTC The PCR product was phosphorad and ligated without enzyme digestion, although we did designed an NheI site in the primers. Since the contruct process contains PCR amplification with Takara LA Taq. The product might have mutations. The desire sequence should be: false false _153_ 0 1596 9 It's complicated false The PCR product is a bit long. So we choose to design long primers for Takara LA Taq. false Zhenyu Shi annotation1974700 1 PstI range1974700 1 19 24 annotation1974702 1 lac promoter range1974702 1 179 208 annotation1974705 1 Mob range1974705 1 1499 2125 annotation1974699 1 SpeI range1974699 1 1 6 annotation1974707 1 lacZalpha reporter range1974707 1 3955 4113 annotation1974710 1 NotI range1974710 1 4164 4171 annotation1974712 1 EcoRI range1974712 1 772 777 annotation1974711 1 EcoRI range1974711 1 25 30 annotation1974706 1 replication protein range1974706 1 2714 3388 annotation1974703 1 chloramphenicol acetyltransferase range1974703 1 301 990 annotation1974708 1 T7 promoter range1974708 1 4121 4139 annotation1974704 1 CAT marker range1974704 1 360 990 annotation1974701 1 T3 promoter range1974701 1 90 109 annotation1974709 1 XbaI range1974709 1 4172 4177 BBa_I713001_sequence 1 actagtggatcccccgggctgcaggaattcgatatcaagcttatcgataccgtcgacctcgagggggggcccggtacccagcttttgttccctttagtgagggttaattgcgcgcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgcatgcataaaaactgttgtaattcattaagcattctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacggaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgataactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgccgatcaacgtctcattttcgccaaaagttggcccagggcttcccggtatcaacagggacaccaggatttatttattctgcgaagtgatcttccgtcacaggtatttattcgaagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcgcccgcgttcctgctggcgctgggcctgtttctggcgctggacttcccgctgttccgtcagcagcttttcgcccacggccttgatgatcgcggcggccttggcctgcatatcccgattcaacggccccagggcgtccagaacgggcttcaggcgctcccgaaggtctcgggccgtctcttgggatgctagctattaattgctagctgacgtactcgaccgccaacacagcgtccttgcgccgcttctctggcagcaactcgcgcagtcggcccatcgcttcatcggtgctgctggccgcccagtgctcgttctctggcgtcctgctggcgtcagcgttgggcgtctcgcgctcgcggtaggcgtgcttgagactggccgccacgttgcccattttcgccagcttcttgcatcgcatgatcgcgtatgccgccatgcctgcccctcccttttggtgtccaaccggctcgacgggggcagcgcaaggcggtgcctccggcgggccactcaatgcttgagtatactcactagactttgcttcgcaaagtcgtgaccgcctacggcggctgcggcgccctacgggcttgctctccgggcttcgccctgcgcggtcgctgcgctcccttgccagcccgtggatatgtggacgatggccgcgagcggccaccggctggctcgcttcgctcggcccgtggacaaccctgctggacaagctgatggacaggctgcgcctgcccacgagcttgaccacagggattgcccaccggctacccagccttcgaccacatacccaccggctccaactgcgcggcctgcggccttgccccatcaatttttttaattttctctggggaaaagcctccggcctgcggcctgcgcgcttcgcttgccggttggacaccaagtggaaggcgggtcaaggctcgcgcagcgaccgcgcagcggcttggccttgacgcgcctggaacgacccaagcctatgcgagtgggggcagtcgaaggcgaagcccgcccgcctgccccccgagcctcacggcggcgagtgcgggggttccaagggggcagcgccaccttgggcaaggccgaaggccgcgcagtcgatcaacaagccccggaggggccactttttgccggagggggagccgcgccgaaggcgtgggggaaccccgcaggggtgcccttctttgggcaccaaagaactagatatagggcgaaatgcgaaagacttaaaaatcaacaacttaaaaaaggggggtacgcaacagctcattgcggcaccccccgcaatagctcattgcgtaggttaaagaaaatctgtaattgactgccacttttacgcaacgcataattgttgtcgcgctgccgaaaagttgcagctgattgcgcatggtgccgcaaccgtgcggcaccctaccgcatggagataagcatggccacgcagtccagagaaatcggcattcaagccaagaacaagcccggtcactgggtgcaaacggaacgcaaagcgcatgaggcgtgggccgggcttattgcgaggaaacccacggcggcaatgctgctgcatcacctcgtggcgcagatgggccaccagaacgccgtggtggtcagccagaagacactttccaagctcatcggacgttctttgcggacggtccaatacgcagtcaaggacttggtggccgagcgctggatctccgtcgtgaagctcaacggccccggcaccgtgtcggcctacgtggtcaatgaccgcgtggcgtggggccagccccgcgaccagttgcgcctgtcggtgttcagtgccgccgtggtggttgatcacgacgaccaggacgaatcgctgttggggcatggcgacctgcgccgcatcccgaccctgtatccgggcgagcagcaactaccgaccggccccggcgaggagccgcccagccagcccggcattccgggcatggaaccagacctgccagccttgaccgaaacggaggaatgggaacggcgcgggcagcagcgcctgccgatgcccgatgagccgtgttttctggacgatggcgagccgttggagccgccgacacgggtcacgctgccgcgccggtagcacttgggttgcgcagcaacccgtaagtgcgctgttccagactatcggctgtagccgcctcgccgccctataccttgtctgcctccccgcgttgcgtcgcggtgcatggagccgggccacctcgacctgaatggaagccggcggcacctcgctaacggattcaccgtttttatcaggctctgggaggcagaataaatgatcatatcgtcaattattacctccacggggagagcctgagcaaactggcctcaggcatttgagaagcacacggtcacactgcttccggtagtcaataaaccggtaaaccagcaatagacataagcggctatttaacgaccctgccctgaaccgacgaccgggtcgaatttgctttcgaatttctgccattcatccgcttattatcacttattcaggcgtagcaccaggcgtttaagggcaccaataactgccttaaaaaaattacgccccgccctgccactcatcgcagtcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaatttccattcgccattcaggctgcgcaactgttgggaagggcgatcggtgcgggcctcttcgctattacgccagctggcgaaagggggatgtgctgcaaggcgattaagttgggtaacgccagggttttcccagtcacgacgttgtaaaacgacggccagtgagcgcgcgtaatacgactcactatagggcgaattggagctccaccgcggtggcggccgctctaga igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z