LOCUS       sequence-451879-        6595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Group B Streptococcus Low Expression Modular Complementation Vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6595)
  AUTHORS   Hillebrand GH, Carlin SC, Giacobe EJ, Stephenson HA, Collins J, 
            Hooven TA
  TITLE     A Cas12a Toolbox for Rapid and Flexible Group B Streptococcus 
            Genomic Editing and CRISPRi.
  JOURNAL   Mol Microbiol. 2025 Nov;124(5):449-461. doi: 10.1111/mmi.70022. Epub
            2025 Sep 13.
  PUBMED    40944362
REFERENCE   2  (bases 1 to 6595)
  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"; doi: "10.1111/mmi.70022";
            journalName: "Mol Microbiol"; date: "2025-11"; volume: "124"; issue:
            "5"; pages: "449-461"
COMMENT     Sequence Label: pGBScomp Low
FEATURES             Location/Qualifiers
     source          1..6595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(559..581)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_RNA        complement(5347..5425)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     rep_origin      complement(5548..6136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5628..5647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6453..6469
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6453..6469
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgaagatct aagcagaact tctatacgtt ctgctttttc ttcattttgt tatttaccaa
       61 tacctttatc tttaatctca gctttttgcc cggtgccatc tactaagggt catttaagtt
      121 gagcatatta gaggaggaaa atcttggaga aatatttgaa gaacccgatt acatggattg
      181 gattagttct tgtggttacg tggtttttaa ctaaaagtag tgaatttttg atttttggtg
      241 tgtgtgtctt gttgttagta tttgctagtc aaagtgatta aatagaattc gaaaagccct
      301 gacaaccctt gttcctaaaa aggaataagc gttcggtcag taaataatag aaataaaaaa
      361 tcagacctaa gactgttgac tttatctaca aggtgtggca taatgtgtgg agaattaaat
      421 taaaaaggga ggccaaatat aatgaaaaat atgaatgaca atgatgttgg atccgtcgac
      481 ctgcagccaa gcttatcgat tctagacgct cagctctttt cttcatttcc ttgtttcctc
      541 ctagcgctct cataccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      601 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      661 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct agcataaaaa
      721 taagaagcct gcatttgcag gcttcttatt tttatggcgc gccgttctga atccttagct
      781 aatggttcaa caggtaacta tgacgaagat agcaccctgg agcaaccagg aatgaattac
      841 tatccctttt atcaagaagc gcacaaaaag aaaaacgaaa tgatacacca atcagtgcaa
      901 aaaaagatat aatgggagat aagacggttc gtgttcgtgc tgacttgcac catatcataa
      961 aaatcgaaac agcaaagaat ggcggaaacg taaaagaagt tatggaaata agacttagaa
     1021 gcaaacttaa gagtgtgttg atagtgcagt atcttaaaat tttgtataat aggaattgaa
     1081 gttaaattag atgctaaaaa tttgtaatta agaaggagtg attacatgaa caaaaatata
     1141 aaatattctc aaaacttttt aacgagtgaa aaagtactca accaaataat aaaacaattg
     1201 aatttaaaag aaaccgatac cgtttacgaa attggaacag gtaaagggca tttaacgacg
     1261 aaactggcta aaataagtaa acaggtaacg tctattgaat tagacagtca tctattcaac
     1321 ttatcgtcag aaaaattaaa actgaatact cgtgtcactt taattcacca agatattcta
     1381 cagtttcaat tccctaacaa acagaggtat aaaattgttg ggagtattcc ttaccattta
     1441 agcacacaaa ttattaaaaa agtggttttt gaaagccatg cgtctgacat ctatctgatt
     1501 gttgaagaag gattctacaa gcgtaccttg gatattcacc gaacactagg gttgctcttg
     1561 cacactcaag tctcgattca gcaattgctt aagctgccag cggaatgctt tcatcctaaa
     1621 ccaaaagtaa acagtgtctt aataaaactt acccgccata ccacagatgt tccagataaa
     1681 tattggaagc tatatacgta ctttgtttca aaatgggtca atcgagaata tcgtcaactg
     1741 tttactaaaa atcagtttca tcaagcaatg aaacacgcca aagtaaacaa tttaagtacc
     1801 gttacttatg agcaagtatt gtctattttt aatagttatc tattatttaa cgggaggaaa
     1861 taattctatg agtcgctttt gtaaatttgg aaagttacac gttactaaag ggaatgtaga
     1921 taaattatta ggtatactac tgacagcttc caaggagcta aagaggtccc tagcgcttag
     1981 aatcgcttta ggaaacacga tccagtccaa taatcgtcga taaaaacttt tgaaaaaggt
     2041 tggtgaaatt acctactttt ggaataatca caaatcacaa gtgattaatc acaaatcaca
     2101 agtgattaat cacttgttta ttaagatatt aaaagctata atttaaataa agcgtgaatt
     2161 ttattacaca aaaagagggg ggagaaactt ggaactagca tttagagaaa gcttaaaaaa
     2221 gatgagaggt accaaatcaa aagaaaaatt ctcccaagaa ttagaaatga gtagatcaaa
     2281 ttattcacga atagaatcag gaaaatcaga tccaaccata aaaacactag aacaaattgc
     2341 aaagttaact aactcaacgc tagtagtgga tttaatccca aatgagccaa cagaaccaga
     2401 accagaaaca gaatcagaac aagtaacatt ggatttagaa atggaagaag aaaaaagcaa
     2461 tgacttcgtg tgaataatgc gcgaaatcgt tgcttatttt ttttaaaagc ggtatactag
     2521 atataacgaa acaacgaact gaatagaaac gaaaaaagag ccatgacaca tttataaaat
     2581 gtttgacgac attttataaa tgcatagccc gataagattg ccaaaccaac gcttatcagt
     2641 tagtcagatg aactcttccc tcgtaagaag ttatttaatt aactttgttt gaagacggta
     2701 tataaccgta ctatcattat atagggaaat cagagagttt tcaagtatct aagctactga
     2761 atttaagaat tgttaagcaa tcaatcggaa atcgtttgat tgcttttttt gtattcattt
     2821 atagaaggtg gagtttgtat gaatcatgat gaatgtaaaa cttatataaa aaatagttta
     2881 ttggagataa gaaaattagc aaatatctat acactagaaa cgtttaagaa agagttagaa
     2941 aagagaaata tctacttaga aacaaaatca gataagtatt tttcttcgga gggggaagat
     3001 tatatatata agttaataga aaataacaaa ataatttatt cgattagtgg aaaaaaattg
     3061 acttataaag gaaaaaaatc tttttcaaaa catgcaatat tgaaacagtt gaatgaaaaa
     3121 gcaaaccaag ttaattaaac aacctatttt ataggattta taggaaagga gaacagctga
     3181 atgaatatcc cttttgttgt agaaactgtg cttcatgacg gcttgttaaa gtacaaattt
     3241 aaaaatagta aaattcgctc aatcactacc aagccaggta aaagcaaagg ggctattttt
     3301 gcgtatcgct caaaatcaag catgattggc ggtcgtggtg ttgttctgac ttccgaggaa
     3361 gcgattcaag aaaatcaaga tacatttaca cattggacac ccaacgttta tcgttatgga
     3421 acgtatgcag acgaaaaccg ttcatacacg aaaggacatt ctgaaaacaa tttaagacaa
     3481 atcaatacct tctttattga ttttgatatt cacacggcaa aagaaactat ttcagcaagc
     3541 gatattttaa caaccgctat tgatttaggt tttatgccta ctatgattat caaatctgat
     3601 aaaggttatc aagcatattt tgttttagaa acgccagtct atgtgacttc aaaatcagaa
     3661 tttaaatctg tcaaagcagc caaaataatt tcgcaaaata tccgagaata ttttggaaag
     3721 tctttgccag ttgatctaac gtgtaatcat tttggtattg ctcgcatacc aagaacggac
     3781 aatgtagaat tttttgatcc taattaccgt tattctttca aagaatggca agattggtct
     3841 ttcaaacaaa cagataataa gggctttact cgttcaagtc taacggtttt aagcggtaca
     3901 gaaggcaaaa aacaagtaga tgaaccctgg tttaatctct tattgcacga aacgaaattt
     3961 tcaggagaaa agggtttaat agggcgtaat aacgtcatgt ttaccctctc tttagcctac
     4021 tttagttcag gctattcaat cgaaacgtgc gaatataata tgtttgagtt taataatcga
     4081 ttagatcaac ccttagaaga aaaagaagta atcaaaattg ttagaagtgc ctattcagaa
     4141 aactatcaag gggctaatag ggaatacatt accattcttt gcaaagcttg ggtatcaagt
     4201 gatttaacca gtaaagattt atttgtccgt caagggtggt ttaaattcaa gaaaaaaaga
     4261 agcgaacgtc aacgtgttca tttgtcagaa tggaaagaag atttaatggc ttatattagc
     4321 gaaaaatcag atgtatacaa gccttattta gtgacgacca aaaaagagat tagagaagtg
     4381 ctaggcattc ctgaacggac attagataaa ttgctgaagg tactgaaggc gaatcaggaa
     4441 attttcttta agattaaacc aggaagaaat ggtggcattc aacttgctag tgttaaatca
     4501 ttgttgctat cgatcattaa agtaaaaaaa gaagaaaaag aaagctatat aaaggcgctg
     4561 acaaattctt ttgacttaga gcatacattc attcaagaga ctttaaacaa gctagcagaa
     4621 cgccctaaaa cggacacaca actcgatttg tttagctatg atacaggctg aaaataaaac
     4681 ccgcactatg ccattacatt tatatctatg atacgtgttt gttttttctt tgctgtttag
     4741 cgaatgatta gcagaaatat acagagtaag attttaatta attattaggg ggagaaggag
     4801 agagtagccc gaaaactttt agttggcttg gactgaacga agtgagggaa aggctactaa
     4861 aacgtcgagg ggcagtgaga gcgaagcgaa cacttgattt tttaattttc tatcttttat
     4921 aggtcattag agtatactta tttgtcctat aaactattta gcagcataat agatttattg
     4981 aataggtcat ttaagttgag catattagag gaggaaaatc ttggagaaat atttgaagaa
     5041 cccgattaca tggattggat tagttcttgt gggtcagtat ttgctagtca aagtgattaa
     5101 atagaattct catgtttgac agcttatcat cggagctccg atgataagct gtcaaacatg
     5161 agaattcccg gggatcctta cgaaatcatc ctgtggagct tagtaggttt agcaagatgg
     5221 cagcgcctaa atgtagaatg ataaaaggat taagagatta atttccctaa aaatgataaa
     5281 acaagcgttt tgaaagcgct tgtttttttg gtttgcagtc agagtagaat agaagtatca
     5341 aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac
     5401 ttgctatgct gttaagaatg gttccaacgc tcactcaaag gcggtaatac ggttatccac
     5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6061 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6121 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6181 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6241 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaagatgtga
     6301 tccgtagcgg tatgcgctcc atcaagaaga gcgacttcgc ggagctggtg aagtacatca
     6361 ccgacgagca aggcaagacc gatcgggccc cctgcaggat aaaaaaattg tagataaatt
     6421 ttataaaata gttttatcta caattttttt atcaggaaac agctatgacc gcggccgctg
     6481 tatccacata aaaataagaa gcctgcattt gcaggcttct tatttttatg gtaccttaag
     6541 acccactttc acatttaagt tgtttttcta atccgcatat gatcaattca aggcc
//
