LOCUS Exported 2696 bp ds-DNA circular SYN 29-MAR-2022 DEFINITION Swapping in small RNA promoter, 23-bp spacer sequence,. ACCESSION . VERSION . KEYWORDS pUCsRNAP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2696) AUTHORS Chua MJ, Collins J TITLE Rapid, Efficient, and Cost-Effective Gene Editing of Enterococcus faecium with CRISPR-Cas12a. JOURNAL Microbiol Spectr. 2022 Feb 2:e0242721. doi: 10.1128/spectrum.02427-21. PUBMED 35107356 REFERENCE 2 (bases 1 to 2696) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 29, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2696 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 112..133 /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=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(606..622) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 650..672 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(710..728) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 796..900 /gene="bla" /label=AmpR promoter CDS 901..1761 /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 complement(1119..1138) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1932..2520 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2421..2440 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2674..2691 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 61 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag 121 ttagctcact cattaggcac cccaggctat tttatgtgag aaatccctaa ataaaaagat 181 gccagtgtgc tggaattcgt cataatcttt aatttgaaaa gatttaaggc ttatttaaat 241 aaaaaatatg agggaagaat tgatataaat ttaattttgt tattgtatta tggtatgtat 301 ggaataaatt taacataaag acagtaataa tgttcttgaa tttagacttt ttatgtgtta 361 tcattaacaa gtatcaaaaa tgacatttaa taaattaata ataattttaa aaatatattt 421 ttgataaaag caatgattaa catggtttga cgtctgagaa gagacgattt tctcaatagg 481 agaaattaag gtgcaaaccc ttatcattcc accataattt ctactcttgt agattcctca 541 aagagcatat ggatatgaat ttctactctt gtagatctcg aggcctgcag acatgcaagc 601 ttggcactgg ccgtcgtttt acaacgtcgc agacaagctg tgaccgtctc cgggagctgc 661 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata 721 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 781 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 841 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 901 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 961 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 1021 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 1081 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 1141 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 1201 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 1261 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 1321 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 1381 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 1441 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 1501 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 1561 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 1621 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 1681 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 1741 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 1801 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 1861 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 1921 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 1981 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2041 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 2101 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2161 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2221 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2281 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2341 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2401 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2461 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2521 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2581 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 2641 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagc //