LOCUS Exported 2526 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pT7tyrgRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2526) AUTHORS Jao LE, Wente SR, Chen W TITLE Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. JOURNAL Proc Natl Acad Sci U S A. 2013 Aug 5. PUBMED 23918387 REFERENCE 2 (bases 1 to 2526) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2526 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 180..202 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 194..211 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 195..211 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_RNA complement(239..314) /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter complement(333..351) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(375..391) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(375..391) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(388..410) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 399..415 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(423..453) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 468..489 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(606..623) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(777..1365) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(857..876) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1536..2396) /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 2159..2178 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2397..2501) /gene="bla" /label=AmpR promoter ORIGIN 1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc 61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg 121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc 181 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggatccgc 241 accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta acttgctatt 301 tctagctcta aaacccccag aagtcctcca gtcctatagt gagtcgtatt aatttcaagc 361 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 421 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 481 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 541 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 601 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 661 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 721 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 781 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 841 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 901 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 961 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 1021 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 1081 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 1141 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 1201 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 1261 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 1321 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 1381 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 1441 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 1501 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 1561 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 1621 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 1681 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 1741 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 1801 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 1861 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 1921 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 1981 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 2041 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 2101 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 2161 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 2221 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 2281 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 2341 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 2401 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 2461 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 2521 ccacct //