LOCUS pBX_The3_sgRNA_b 5511 bp ds-DNA circular 01-OCT-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBX The3 BbsI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5522) AUTHORS MIT TITLE Direct Submission JOURNAL Exported Tuesday, Apr 19, 2016 from SnapGene 3.1.0 http://www.snapgene.com COMMENT COMMENT ApEinfo:methylated:1 FEATURES Location/Qualifiers CDS 458..1252 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR P VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL S SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH Q GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI A LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" /locus_tag="aminoglycoside phosphotransferase from Tn5" /label=aminoglycoside phosphotransferase from Tn5 /ApEinfo_fwdcolor=#e9d024 /ApEinfo_revcolor=#e9d024 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 terminator 1600..1646 /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" /locus_tag="rrnB T1 terminator" /label=rrnB T1 terminator /ApEinfo_fwdcolor=#9d1b1c /ApEinfo_revcolor=#9d1b1c /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_feature 1979..2064 /note="The3 sgRNA" /locus_tag="The3 sgRNA" /label=The3 sgRNA /ApEinfo_fwdcolor=#7eff74 /ApEinfo_revcolor=#7eff74 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 terminator 2070..2113 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" /locus_tag="Escherichia coli rrnB" /label=Escherichia coli rrnB /ApEinfo_fwdcolor=#9d1b1c /ApEinfo_revcolor=#9d1b1c /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer_bind complement(2256..2272) /note="SK primer" /note="common sequencing primer, one of multiple similar variants" /locus_tag="SK primer" /label=SK primer /ApEinfo_fwdcolor=#14c0bd /ApEinfo_revcolor=#4ec02b /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 promoter complement(2305..2323) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" /locus_tag="T7 promoter" /label=T7 promoter /ApEinfo_fwdcolor=#346ee0 /ApEinfo_revcolor=#346ee0 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer_bind complement(2333..2349) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" /locus_tag="M13 fwd" /label=M13 fwd /ApEinfo_fwdcolor=#14c0bd /ApEinfo_revcolor=#4ec02b /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 CDS complement(3057..3719) /codon_start=1 /product="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" /note="pBBR1 Rep" /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLL H HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYV V NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGE E PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR" /locus_tag="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" /label=replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica /ApEinfo_fwdcolor=#e9d024 /ApEinfo_revcolor=#e9d024 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 rep_origin 3720..4489 /note="pBBR1 oriV" /note="replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication" /locus_tag="pBBR1 oriV" /label=pBBR1 oriV /ApEinfo_fwdcolor=#999999 /ApEinfo_revcolor=#999999 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_feature 1661..1958 /label=pxyl /ApEinfo_fwdcolor=#808000 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_feature complement(1959..1978) /label=full_guide1 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=#00ffff /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ORIGIN 1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc 61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg 121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg 181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag 241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt 301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa 361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc 421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca 1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta 1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga 1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatata agctttcagt 1561 cggcgcgaat gctccacgcc gcgcccagcg gcgcggatcc aaataaaacg aaaggctcag 1621 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgc tgcagccagc cgtggtcggg 1681 cagcaggtag aaggcgccct cgtcctgatc ctcgcccgaa acctccagcc cccggtcgat 1741 ggcttcgacg acatagccgg ccgcgcggca ggtgtcggtg agcgcggcca gcagggcggc 1801 ttcctggtca ggggtcaggt cggtcatggg caagaggtcc aggtcgtggt ttgtcggcgg 1861 cttctagcat ggaccgcccg cgcccgtgag gccgaggatt tcgcgctggt cagacaacct 1921 acttgccgtc cccacatgtt agcgctacca agtgccgaCC AGGATCAGGC GCTTCATGgt 1981 tttagagctg tgaaaacagc gagttaaaat aaggcttagt ccgtactcaa cttgaaaagg 2041 tggcaccgat tcggtgtttt ttttttttga taaaacgaaa ggcccagtct ttcgactgag 2101 cctttcgttt tataaacggg tcttgagggg ttttttaagc ttctgcagct ggatccatgg 2161 atatctagat ttccgcaaaa taaaaaaggg gacctctagg gtccccaatt aattagctaa 2221 agccgctagc ccatgggaat tcctgcagcc cgggggatcc actagttcta gagcggccgc 2281 caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg 2341 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag 2401 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc 2461 aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt 2521 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgtac tgcgatgagt 2581 ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggtgcta 2641 cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg 2701 gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg gtttaccggt 2761 ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg ccagtttgct 2821 caggctctcc ccgtggaggt aataattgac gatatgatca tttattctgc ctcccagagc 2881 ctgataaaaa cggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg 2941 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgaggc 3001 ggctacagcc gatagtctgg aacagcgcac ttacgggttg ctgcgcaacc caagtgctac 3061 cggcgcggca gcgtgacccg tgtcggcggc tccaacggct cgccatcgtc cagaaaacac 3121 ggctcatcgg gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc 3181 aaggctggca ggtctggttc catgcccgga atgccgggct ggctgggcgg ctcctcgccg 3241 gggccggtcg gtagttgctg ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca 3301 tgccccaaca gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc actgaacacc 3361 gacaggcgca actggtcgcg gggctggccc cacgccacgc ggtcattgac cacgtaggcc 3421 gacacggtgc cggggccgtt gagcttcacg acggagatcc agcgctcggc caccaagtcc 3481 ttgactgcgt attggaccgt ccgcaaagaa cgtccgatga gcttggaaag tgttttctgg 3541 ctgaccacca cggcgttctg gtggcccatc tgcgccacga ggtgatgcag cagcattgcc 3601 gccgtgggtt tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc 3661 acccagtgac cgggcttgtt cttggcttga atgccgattt ctctggactg cgtggccatg 3721 cttatctcca tgcggtaggg tgccgcacgg ttgcggcacc atgcgcaatc agctgcaact 3781 tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg gcagtcaatt acagattttc 3841 tttaacctac gcaatgagct attgcggggg gtgccgcaat gagctgttgc gtacccccct 3901 tttttaagtt gttgattttt aagtctttcg catttcgccc tatatctagt tctttggtgc 3961 ccaaagaagg gcacccctgc ggggttcccc cacgccttcg gcgcggctcc ccctccggca 4021 aaaagtggcc cctccggggc ttgttgatcg actgcgcggc cttcggcctt gcccaaggtg 4081 gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg ctcggggggc aggcgggcgg 4141 gcttcgcctt cgactgcccc cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc 4201 aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg tccaaccggc 4261 aagcgaagcg cgcaggccgc aggccggagg cttttcccca gagaaaatta aaaaaattga 4321 tggggcaagg ccgcaggccg cgcagttgga gccggtgggt atgtggtcga aggctgggta 4381 gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt 4441 ccagcagggt tgtccacggg ccgagcgaag cgagccagcc ggtggccgct cgcggccatc 4501 gtccacatat ccacgggctg gcaagggagc gcagcgaccg cgcagggcga agcccggaga 4561 gcaagcccgt agggcgccgc agccgccgta ggcggtcacg actttgcgaa gcaaagtcta 4621 gtgagtatac tcaagcattg agtggcccgc cggaggcacc gccttgcgct gcccccgtcg 4681 agccggttgg acaccaaaag ggaggggcag gcatggcggc atacgcgatc atgcgatgca 4741 agaagctggc gaaaatgggc aacgtggcgg ccagtctcaa gcacgcctac cgcgagcgcg 4801 agacgcccaa cgctgacgcc agcaggacgc cagagaacga gcactgggcg gccagcagca 4861 ccgatgaagc gatgggccga ctgcgcgagt tgctgccaga gaagcggcgc aaggacgctg 4921 tgttggcggt cgagtacgtc atgacggcca gcccggaatg gtggaagtcg gccagccaag 4981 aacagcaggc ggcgttcttc gagaaggcgc acaagtggct ggcggacaag tacggggcgg 5041 atcgcatcgt gacggccagc atccaccgtg acgaaaccag cccgcacatg accgcgttcg 5101 tggtgccgct gacgcaggac ggcaggctgt cggccaagga gttcatcggc aacaaagcgc 5161 agatgacccg cgaccagacc acgtttgcgg ccgctgtggc cgatctaggg ctgcaacggg 5221 gcatcgaggg cagcaaggca cgtcacacgc gcattcaggc gttctacgag gccctggagc 5281 ggccaccagt gggccacgtc accatcagcc cgcaagcggt cgagccacgc gcctatgcac 5341 cgcagggatt ggccgaaaag ctgggaatct caaagcgcgt tgagacgccg gaagccgtgg 5401 ccgaccggct gacaaaagcg gttcggcagg ggtatgagcc tgccctacag gccgccgcag 5461 gagcgcgtga gatgcgcaag aaggccgatc aagcccaaga gacggcccga g //