BBa_K823054 1 Sporovecto <b>Sporo</b>vector 2012-09-22T11:00:00Z 2015-05-08T01:13:30Z Assembly of several PCRs. For N-terminal fusion proteins with cgeA (display on B. subtilis spores).=='''Sporo'''vector== Our '''Sporo'''vector is specially designed to give you the opportunity to easily create Sporobeads with any proteins you can think of. It is available as BioBrick in pSB1C3 and can be cloned in any of our empty ''B. subtilis'' vectors. We also offer it ???precloned??? in pSB_Bs4S which in this version lacks ''NgoM''IV restriction sites. [[File:Sporovectorcloning.png|600px]] The '''Sporo'''vector was created by ligation of three cut PCR-products. It is designed to allow N-terminal fusion of genes in Freiburg standard ([http://dspace.mit.edu/bitstream/handle/1721.1/45140/BBF_RFC%2025.pdf?sequence=1 RCF25]). Therefore, the gene to be inserted must be cut with ''Xba''I and ''Age''I and the vector must be digested with ''Xba''I and ''NgoM''IV. By this digest, the RFP cassette will be removed from this vector to allow easy selection for the integration of the new insert. After ligation, there is a scar of six nucleotides between both genes but no restriction site left. The gene fusion is controlled by the P_''cot''YZ-promoter which was the strongest one in our promoter evaluations. The C-terminal gene part is ''cge''A, one of the two known spore crust proteins. The vector is cloned but has not been tested with an insert, yet. We are working on further '''Sporo'''vectors to offer also fusion with ''cot''Z as well as C-terminal fusions. false false _1081_ 0 6378 9 It's complicated true For N-terminal fusion proteins with cgeA (display on B. subtilis spores) false Jara Radeck annotation2195356 1 NgoMIV range2195356 1 2035 2040 annotation2195354 1 XbaI range2195354 1 959 964 annotation2195351 1 thrC range2195351 1 247 756 annotation2195355 1 RFP range2195355 1 1192 1872 annotation2195352 1 EcoRI range2195352 1 766 772 annotation2195357 1 cgeA range2195357 1 2038 2439 annotation2195364 1 bla range2195364 1 5243 6103 annotation2195360 1 PstI range2195360 1 2543 2548 annotation2195361 1 spec range2195361 1 2738 3412 annotation2195363 1 thrB range2195363 1 3980 4390 annotation2195359 1 B0014 range2195359 1 2448 2542 annotation2195353 1 PcotYZ range2195353 1 773 958 annotation2195358 1 AgeI range2195358 1 2431 2436 annotation2195350 1 hom range2195350 1 1 247 annotation2195365 1 hom range2195365 1 6258 6313 annotation2195362 1 thrC range2195362 1 3555 3983 BBa_K823054_sequence 1 aattcatgtaaaagatgaggttggttcattctcgaaaattacatctgtgttctcagagcggggcgtgacgtttgaaaaaatccttcagctgccaattaaaggccatgatgagttagctgaaatcgtaattgtcacacatcatacatcagaagctgatttcagtgatatcctgcaaaacctaaatgatttggaagtcgttcaagaagtcaaaagcacatatcgtgtagaagggaacggttggagctaatgtggaaaggacttatccatcaatataaagaatttttacctgtaacagatcaaacaccggcgctaactttacatgaaggaaacacacctcttattcacctgccgaagctgtctgagcagctcggaattgagcttcatgtcaaaacggaaggcgtcaatcctacgggatcatttaaagatcgcggaatggttatggctgtggcaaaggcaaaagaagaaggcaatgacacgattatgtgcgcgtcaacaggtaacacttccgctgctgcggcagcatatgcagcccgtgctaacatgaaatgcattgtcatcatcccgaacggaaaaattgcatttggaaaactcgctcaagctgtcatgtacggagccgagattatcgcaattgacggaaactttgacgatgcgcttaaaattgtccgttccatctgtgagaaatcaccgattgcccttgtcaactcagtcaacccttaccgcattgaaggccaaaaaactgctgccttcggatcctagaagcttatcgaattcgacagcaacaaatacactcgtagccatcctagttatcactcttgtcctctaggacctaaaagcagagctaaaaacgctctgctttttcttattttccaagcatatgatgaatatatagacgttcacccacaccaagtggggcacgggtacatatgttgttaaggactaaagtcaaataccctataattctagagcaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacatactagagaaagaggagaaatactagatggcttcctccgaagacgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagacggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgatggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagacggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaacctgttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagactacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactcctccggagcttaataacgctgatagtgctagtgtagatcgctactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatagccggcgaaaatgcacagttaaaaaaggatttaattaaagccgttttaagccctttatttccaactgcaacagaaggaggagaaaatatggatagtaatcttaaagccttgcttgatgctgccatcgatcaaaaagtagatgaaagtgaaacggttacggcagaatctattttagacccttctcttccggcaagatggatttttgccagaattacgccaggcactacaatcagcattgtgactgattcaggtgacatgatcggaccagttgttttcgttgctttcgatcaggttcacgggatcgtatttgtaacacaggaaagctccgtcactccggcaggccaagctacaacattaattgatgtagacaaagtagaaagcgttacgttcttttcaaccggttaatactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattatttctgcaggataaaaaatttagaagccaatgaaatctataaataaactaaattaagtttatttaattaacaactatggatataaaataggtactaatcaaaatagtgaggaggatatatttgaatacatacgaacaaattaataaagtgaaaaaaatacttcggaaacatttaaaaaataaccttattggtacttacatgtttggatcaggagttgagagtggactaaaaccaaatagtgatcttgactttttagtcgtcgtatctgaaccattgacagatcaaagtaaagaaatacttatacaaaaaattagacctatttcaaaaaaaataggagataaaagcaacttacgatatattgaattaacaattattattcagcaagaaatggtaccgtggaatcatcctcccaaacaagaatttatttatggagaatggttacaagagctttatgaacaaggatacattcctcagaaggaattaaattcagatttaaccataatgctttaccaagcaaaacgaaaaaataaaagaatatacggaaattatgacttagaggaattactacctgatattccattttctgatgtgagaagagccattatggattcgtcagaggaattaatagataattatcaggatgatgaaaccaactctatattaactttatgccgtatgattttaactatggacacgggtaaaatcataccaaaagatattgcgggaaatgcagtggctgaatcttctccattagaacatagggagagaattttgttagcagttcgtagttatcttggagagaatattgaatggactaatgaaaatgtaaatttaactataaactatttaaataacagattaaaaaaattataaaaaaattgaaaaaatggtggaaacacttttttcaatttttttgttttattatttaatatttgggaaatattcattctaattggtaatcagattttagaaaacaataaacccttgcatagggggatctcgaggccttccgaaaatgcgcggctttgaagctgagggtgcggcggcaatcgtgcgcaatgaagtgattgaaaatccggaaacaatagcgacagccattcgtatcggaaacccggcaagctgggacaaagctgtaaaggcagccgaggaatccaatgggaaaattgacgaagtcactgatgatgaaatccttcacgcatatcagctgatcgcccgtgtagaaggcgtgtttgcagaaccaggttcttgcgcgtctatcgcaggagtgctgaaacaggtgaaatccggagaaattccgaaaggcagcaaggtcgtagctgtgttaacaggaaacggactgaaagatccgaacacagcggtcgacatttcagaaatcaagcctgtcacattgccgactgatgaagacagcatccttgaatatgtaaaaggagcggcccgtgtatgaacgaagccgacatgctgttctctgtcactgttcccggaagcacagctaacctaggccccggctttgattcagtcggaatggcgctcagcagatatttgaagctgaccgtctttgaaagcgacaaatggtcttttgaggctgaaacagaaacagtcgccggaattaattcgagctcctttaacttcctcaccagtagttgtatcggtaccataagtagaagcagcaacccaagtagctttaccagcatccggttcaaccagcatagtaagaatcttactggacatcggcagttcttcgaacagtgcgccaactaccagctctttctacagttcattcagggcaccggagaacctgcgtgcaatccatcttgttcaatcatgcgaaacgatcctcatcctgtctcttgatccatggattacgcgcgaccttcagcatcaccggcatgtccccctggcggacgggaagtatccagctcgaggtcgggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttgatagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctgctggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaacacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggccctttcgtcttcaagaatt igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z