LOCUS Exported 4379 bp ds-DNA circular SYN 17-OCT-2014 DEFINITION . ACCESSION . VERSION . KEYWORDS piG0067_logic_bxb1_BUFFER-sfGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4379) AUTHORS Daniel Gerngross TITLE Direct Submission JOURNAL Exported 17 Oct 2014 from SnapGene 2.5.0 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4379 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 1..126 /note="T0" terminator 27..121 /note="lambda t0 terminator" /note="transcription terminator from phage lambda" promoter 165..199 /note="J23100" misc_feature 200..249 /note="attB Bxb1" misc_feature 224..225 /note="conserved" terminator 250..378 /note="B0015" /note="directional double term" terminator 258..329 /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 345..372 /note="T7Te terminator" /note="phage T7 early transcription terminator" misc_feature complement(379..436) /note="attP Bxb1" misc_feature 407..408 /note="conserved" misc_feature 437..456 /note="35" /note="35 nt upstream of ATG from bla gene from pUC18 (bla RBS) --> introduction of XbaI site for library cloning" CDS 472..1197 /codon_start=1 /note="sfGFP" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYKRP*" CDS 1192..1194 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /note="mRFP1" /translation="" terminator 1209..1295 /gene="Escherichia coli rrnB" /note="T1" /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin 1410..2181 /note="pBBR1 oriV" /note="replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication" rep_origin 2059..2844 /note="pBBR1" CDS 2182..2844 /codon_start=1 /product="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" /note="pBBR1 Rep" /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR" oriT 2855..2963 /note="incP origin of transfer" CDS complement(3109..4299) /note="TetR" ORIGIN 1 aaatcgtaat tattggggac ccctggattc tcaccaataa aaaacgcccg gcggcaaccg 61 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt 121 ccaagactag tcgccagggt tttcccagtc acgacgcggc cgcattgacg gctagctcag 181 tcctaggtac agtgctagct cggccggctt gtcgacgacg gcggtctccg tcgtcaggat 241 catccgggcc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt 301 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg 361 ggcctttctg cgtttatagt cggggtttgt accgtacacc actgagaccg cggtggttga 421 ccagacaaac cacgacgata aatgcttcaa taatatgagg aggtgatcta gatgagcaaa 481 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat 541 gggcacaaat tttctgtccg tggagagggt gaaggtgatg ctacaaacgg aaaactcacc 601 cttaaattta tttgcactac tggaaaacta cctgttccgt ggccaacact tgtcactact 661 ctgacctatg gtgttcaatg cttttcccgt tatccggatc acatgaaacg gcatgacttt 721 ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt caaagatgac 781 gggacctaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt taatcgtatc 841 gagttaaagg gtattgattt taaagaagat ggaaacattc ttggacacaa actcgagtac 901 aactttaact cacacaatgt atacatcacg gcagacaaac aaaagaatgg aatcaaagct 961 aacttcaaaa ttcgccacaa cgttgaagat ggttccgttc aactagcaga ccattatcaa 1021 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtcgaca 1081 caatctgtcc tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct tcttgagttt 1141 gtaactgctg ctgggattac acatggcatg gatgagctct acaaaaggcc ttaataaatt 1201 aaaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt 1261 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagaca gctgggcgcg 1321 ccccccctac gggcttgctc tccgggcttc gccctgcgcg gtcgctgcgc tcccttgcca 1381 gcccgtggat atgtggacga tggccgcgag cggccaccgg ctggctcgct tcgctcggcc 1441 cgtggacaac cctgctggac aagctgatgg acaggctgcg cctgcccacg agcttgacca 1501 cagggattgc ccaccggcta cccagccttc gaccacatac ccaccggctc caactgcgcg 1561 gcctgcggcc ttgccccatc aattttttta attttctctg gggaaaagcc tccggcctgc 1621 ggcctgcgcg cttcgcttgc cggttggaca ccaagtggaa ggcgggtcaa ggctcgcgca 1681 gcgaccgcgc agcggcttgg ccttgacgcg cctggaacga cccaagccta tgcgagtggg 1741 ggcagtcgaa gggcgaagcc cgcccgcctg ccccccgagc ctcacggcgg cgagtgcggg 1801 ggttccaagg gggcagcgcc accttgggca aggccgaagg ccgcgcagtc gatcaacaag 1861 ccccggaggg gccacttttt gccggagggg gagccgcgcc gaaggcgtgg gggaaccccg 1921 caggggtgcc cttctttggg caccaaagaa ctagatatag ggcgaaatgc gaaagactta 1981 aaaatcaaca acttaaaaaa ggggggtacg caacagctca ttgcggcacc ccccgcaata 2041 gctcattgcg taggttaaag aaaatctgta attgactgcc acttttacgc aacgcataat 2101 tgttgtcgcg ctgccgaaaa gttgcagctg attgcgcatg gtgccgcaac cgtgcggcac 2161 ccctaccgca tggagataag catggccacg cagtccagag aaatcggcat tcaagccaag 2221 aacaagcccg gtcactgggt gcaaacggaa cgcaaagcgc atgaggcgtg ggccgggctt 2281 attgcgagga aacccacggc ggcaatgctg ctgcatcacc tcgtggcgca gatgggccac 2341 cagaacgccg tggtggtcag ccagaagaca ctttccaagc tcatcggacg ttctttgcgg 2401 acggtccaat acgcagtcaa ggacttggtg gccgagcgct ggatctccgt cgtgaagctc 2461 aacggccccg gcaccgtgtc ggcctacgtg gtcaatgacc gcgtggcgtg gggccagccc 2521 cgcgaccagt tgcgcctgtc ggtgttcagt gccgccgtgg tggttgatca cgacgaccag 2581 gacgaatcgc tgttggggca tggcgacctg cgccgcatcc cgaccctgta tccgggcgag 2641 cagcaactac cgaccggccc cggcgaggag ccgcccagcc agcccggcat tccgggcatg 2701 gaaccagacc tgccagcctt gaccgaaacg gaggaatggg aacggcgcgg gcagcagcgc 2761 ctgccgatgc ccgatgagcc gtgttttctg gacgatggcg agccgttgga gccgccgaca 2821 cgggtaacgc tgccgcgccg gtagggccgg cctacggcca gcctcgcaga gcaggattcc 2881 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg 2941 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac 3001 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg 3061 ctataatgac cccgaagcag ggttatgcag cggaaaagga caattgtctc aggtcgaggt 3121 ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc 3181 tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat 3241 cagcggtcca gtgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc 3301 ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc 3361 gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg cgaacgccag 3421 caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa 3481 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac 3541 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac 3601 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc 3661 ggccacaatg gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa 3721 gggcatcgga cggcgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag 3781 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgcg tgcagggaga tggcgcccaa 3841 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 3901 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 3961 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagagaatcc acaggacggg 4021 tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga gcaggactgg 4081 gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa attgcatcaa 4141 cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat ggacgatatc 4201 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac 4261 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta 4321 gcaatttaac tgtgataaac taccgcatta cagtttatcg atgataagct gtcaaattt //