Plasmid
Part:BBa_I713001:Design
Designed by: Zhenyu Shi Group: iGEM07_Tsinghua (2007-10-25)
A pBBR1MCS plasmid without the mob gene.
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 25
Illegal EcoRI site found at 772
Illegal XbaI site found at 4172
Illegal SpeI site found at 1
Illegal PstI site found at 19 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 25
Illegal EcoRI site found at 772
Illegal NheI site found at 1484
Illegal NheI site found at 1498
Illegal SpeI site found at 1
Illegal PstI site found at 19
Illegal NotI site found at 4164 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 25
Illegal EcoRI site found at 772
Illegal BamHI site found at 7
Illegal XhoI site found at 58 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 25
Illegal EcoRI site found at 772
Illegal XbaI site found at 4172
Illegal SpeI site found at 1
Illegal PstI site found at 19 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 25
Illegal EcoRI site found at 772
Illegal XbaI site found at 4172
Illegal SpeI site found at 1
Illegal PstI site found at 19
Illegal NgoMIV site found at 3525
Illegal AgeI site found at 3685 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1461
Design Notes
The PCR product is a bit long. So we choose to design long primers for Takara LA Taq.
- RS 18:18, 4 September 2008 (UTC): The sequence for this part has been reentered to begin with SpeI in keeping with the BioBrick plasmid convention. Feature annotations have been added.
Source
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
//