LOCUS       sequence-358079-        2696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Swapping in small RNA promoter, 23-bp spacer sequence,.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microbiol 
            Spectr. 2022 Feb 2:e0242721. doi: 10.1128/spectrum.02427-21."
COMMENT     Sequence Label: pUCsRNAP
FEATURES             Location/Qualifiers
     source          1..2696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(901..969,970..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
//
