LOCUS Exported 2302 bp ds-DNA circular SYN 05-JUN-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2302) AUTHORS AVA TITLE Direct Submission JOURNAL Exported Wednesday, Nov 6, 2019 from SnapGene 5.0.0 https://www.snapgene.com FEATURES Location/Qualifiers source 1..2302 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 24..46 /label=AVA037 promoter 82..330 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 189..208 /label=KZ396 primer_bind 253..272 /label=LKO.1 5' primer_bind complement(312..353) /label=AVA091 primer_bind 326..374 /label=AVA092 misc_feature 331..350 /label=spacer misc_RNA 351..426 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 427..440 /label=RT template misc_feature 436 /label=G to T misc_feature 441..455 /label=annealing primer_bind complement(478..501) /label=AVA038 rep_origin complement(553..1141) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(716..735) /label=KZ397 primer_bind 1216..1235 /label=ACK562 CDS complement(1312..2172) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1718..1737 /label=ACK563 promoter complement(2173..2277) /gene="bla" /label=AmpR promoter promoter complement(2214..2242) /label=amp prom ORIGIN 1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc 61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata 121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta 181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta 241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct 301 ttatatatct tgtggaaagg acgaaacacc gtcatcttag tcattacctg gttttagagc 361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt 421 cggtgcaacg aacacatcag gtaatgacta agatgttttt ttgggccgct cgaggtacct 481 ctctacatat gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 541 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 601 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 661 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 721 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 781 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 841 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 901 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 961 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 1021 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 1081 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 1141 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 1201 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 1261 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 1321 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 1381 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 1441 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 1501 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 1561 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 1621 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 1681 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 1741 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 1801 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 1861 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 1921 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 1981 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 2041 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 2101 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 2161 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 2221 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 2281 atttccccga aaagtgccac ct //