Composite

Part:BBa_K243012:Design

Designed by: Freiburg Bioware09   Group: iGEM09_Freiburg_bioware   (2009-10-14)
Revision as of 16:13, 16 October 2009 by Timo (Talk | contribs) (→‎Source)

Strep-DigA-Middle Linker-Fok_i


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 278
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The choice of the linker, lipocalin tag and purification tag allows us serval different combinations.


Source

Combined the parts by serial cloning steps.

LOCUS Strep+Dig+middel 3625 bp DNA circular SYN 13-OCT-2009 DEFINITION Ligation of Strep+Dig+middellinker+FokI into pMA2_NgoMIV_SpeI ACCESSION Strep+Dig+middel KEYWORDS . SOURCE Unknown. 

 ORGANISM  Unknown
           Unclassified.

REFERENCE 1 (bases 1 to 3625) 

 AUTHORS   Self
 JOURNAL   Unpublished.

COMMENT SECID/File created by SciEd Central, Scientific & Educational Software COMMENT SECNOTES|Vector molecule: pMA2_NgoMIV_SpeI 

              Fragment ends: SpeI and NgoMIV
              Fragment size: 2412
           Insert molecule:  Strep+Dig+middellinker+FokI
              Fragment ends: NgoMIV and SpeI
              Fragment size: 1213

FEATURES Location/Qualifiers 

    CDS             129..989
                    /gene="AmpR"
                    /SECDrawAs="Gene"
    CDS             2008..3222
                    /gene="ORF-1"
                    /SECDrawAs="Gene"
    CDS             2014..2041
                    /gene="'Strep-Tag'"
                    /SECDrawAs="Gene"
    CDS             2044..2599
                    /gene="'Dig'"
                    /SECDrawAs="Gene"
    CDS             2602..2628
                    /gene="'middelLinker38"
                    /SECDrawAs="Gene"
    CDS             2632..3219
                    /gene="'FokI"
                    /SECDrawAs="Gene"

ORIGIN 

       1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     841 gtggttctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
    1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
    1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
    1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
    1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
    1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
    1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
    1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
    1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
    1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
    1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
    1801 ctcattaggc accccaggct ttacccgaac gaccgagcgc agcgagtcag tgagcgagga
    1861 agcggagagc gcccaatacg caaggaaaca gctatgacca tgttaatgca gctggcacga
    1921 caggtttccc gactggaaag cgggcagtga gcggaaggcc catgaggcca gttaattaag
    1981 aggtaccgaa ttcgcggccg cttctagatg gccggctgga gccatccgca gtttgaaaaa
    2041 accggcgacg tgtaccacga cggcgcctgc cccgaagtga agcccgtgga caacttcgac
    2101 tggtcccagt accacggcaa gtggtggcag gtggccgctt atcccgacca catcaccaag
    2161 tacggcaagt gcggctgggc cgagtacacc cccgagggca agagcgtgaa ggtgtcccgg
    2221 tacagcgtga tccacggcaa agagtacttc agcgagggca ccgcctaccc tgtgggcgac
    2281 agcaagatcg gcaagatcta ccacagctac accatcggcg gcgtgaccca ggtgggcgtg
    2341 agcaacgtgc tgtccaccga caacaagaac tacatcatcg gctacttttg cagatacgac
    2401 gaggacaaga agggccactg ggacgccgtg tgggtgctgt cccggtccat ggtgctgacc
    2461 ggcgaggcca agaccgccgt ggagaactac ctgatcggca gccccgtggt ggacagccag
    2521 aaactggtgt acagcgactt ctccgaggcc gcctgcaaag tgaacaacag caactggtcc
    2581 cacccccagt tcgaaaagac cggcggtggt tctggtggtg gttctggtac cggcaaatct
    2641 gaactggagg agaaaaaatc cgagctgcgc cacaaactga aatatgtgcc tcacgagtat
    2701 atcgaactga tcgagatcgc ccgtaatagt acccaagacc gtatcctgga aatgaaagtg
    2761 atggagttct tcatgaaagt ctatggctat cgtggcaaac atctgggtgg tagccgtaaa
    2821 ccagcaggtg ccatttatac cgttggttcc ccgatcgatt atggcgttat cgtggccaca
    2881 aaagcgtatt ctggcggtta taatctgccg attggtcagg ctgatgagat ggaacgttat
    2941 gtggaagaga atcagacccg taacaaacat ctgaacccga acgaatggtg gaaagtgtat
    3001 ccgtcaagtg tcaccgagtt caaatttctg ttcgtgagcg gccactttaa aggcaactat
    3061 aaagcccagc tgactcgtct gaaccatatc accaatagca atggggcagt gctgagtgtt
    3121 gaggaactgc tgatcggtgg agaaatgatc aaagcaggca ccctgactct ggaagaagtt
    3181 cgccgtaaat tcaacaatgg cgagatcaat tttaccggtt aatactagta gcggccgctg
    3241 cagagctcat ggcgcgccta ggccttgacg gccttccgcc aattcgccct atagtgagtc
    3301 gtattacgtc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
    3361 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
    3421 aggcccgcac cgaaacgccc ttcccaacag ttgcgcagcc tgaatggcga atgggagcgc
    3481 cctgtagcgg ccactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
    3541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
    3601 caaaatatta acgcttacaa tttag

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References