Coding

Part:BBa_K806001:Design

Designed by: Chang Ye   Group: iGEM12_SYSU-China   (2012-09-24)


iciA-inhibitor of chromosomal initiation of replication


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 256
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. Add restriction enzyme in the front term of PCR primers 2. Using genomic DNA extraction as a template of PCR we have better effect


Source

We clone this gene from E.coli strain BL21 with primer iciA-F and iciA-R

iciA-F sequence:GCTCTAGAGCATGAAACGCCCGGACTACAGAAC iciA-R sequence:GGACTAGTCCTTAATCCTGACGAAGGACTTTGTGAC iciA sequence:(GenBank: CP001509.3) 

      1 atgaaacgcc cggactacag aacattacag gcactggatg cggtgatacg tgaacgagga

     61 tttgagcgcg cggcacaaaa gctgtgcatt acacaatcag ccgtctcaca gcgcattaag

    121 caactggaaa atatgttcgg gcagccgctg ttggtgcgta ccgtaccgcc gcgcccgacg

    181 gaacaagggc aaaaactgct ggcactgctg cgccaggtgg agttgctgga agaagagtgg

    241 ctgggcgatg aacaaaccgg ttcgactccg ctgctgcttt cactggcggt caacgccgac

    301 agtctggcga cgtggttgct tcctgcactg gctcctgtgt tggctgattc gcctatccgc

    361 ctcaacttgc aggtagaaga tgaaacccgc actcaggaac gtctgcgccg cggcgaagtg

    421 gtcggcgcgg tgagtattca acatcaggcg ctgccgagtt gtcttgtcga taaacttggt

    481 gcgctcgact atctgttcgt cagctcaaaa ccctttgccg aaaaatattt ccctaacggc

    541 gtaacgcgtt cggcattact gaaagcgcca gtggtcgcgt ttgaccatct tgacgatatg

    601 caccaggcct ttttgcagca aaacttcgat ctgcctccag gcagcgtgcc ctgccatatc

    661 gttaattctt cagaagcgtt cgtacaactt gctcgccagg gcaccacctg ctgtatgatc

    721 ccgcacctgc aaatcgagaa agagctggcc agcggtgaac tgattgactt aacgcctggg

    781 ctatttcaac gacggatgct ctactggcac cgctttgctc ctgaaagccg catgatgcgt

    841 aaagtcactg atgcgttact cgattatggt cacaaagtcc ttcgtcagga ttaa

References