LOCUS       sequence-274098-        5156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli-Lactobacilli shuttle vector for recombineering template 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5156)
  AUTHORS   Leenay RT, Vento JM, Shah M, Martino ME, Leulier F, Beisel CL
  TITLE     Genome Editing with CRISPR-Cas9 in Lactobacillus plantarum Revealed 
            That Editing Outcomes Can Vary Across Strains and Between Methods.
  JOURNAL   Biotechnol J. 2019 Mar;14(3):e1700583. doi: 10.1002/biot.201700583. 
            Epub 2018 Sep 20.
  PUBMED    30156038
REFERENCE   2  (bases 1 to 5156)
  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: "Biotechnol 
            J."; date: "2019-03"; volume: "14(3):e1700583. doi"; pages: " 
            10.1002/biot.201700583. Epub 2018 Sep 20"
COMMENT     Sequence Label: pCB591
FEATURES             Location/Qualifiers
     source          1..5156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(641..652)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2906..2997
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2998..3066,3067..3858)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3216..3235)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3900..4355
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3987..4006)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4466..5054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4955..4974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgcctttg agtgagcccg atgaagctga cagctaaggt accactagtt ctagagagct
       61 caagctttct ttgaaccaaa attagaaaac caaggcttga aacgttcaat tgaaatggca
      121 attaaacaaa ttacagcacg tgttgctttg attgatagcc aaaaagcagc agttgataaa
      181 gcaattactg atattgctga aaaattgtaa tttataaata aaaatcacct tttagaggtg
      241 gtttttttat ttataaatta ttcgtttgat ttcgctttcg atagaacaat caaagcgaga
      301 ataaggaaga taaatcccat aagggcggga gcagaatgtc cgagactaat gtaaatttgt
      361 ccaccaatta aaggaccgat aacgcgagct tctcgagtgc atattttcgg caatcttctc
      421 aatgagatgc tcttcagcat gttcaatgat gtcgattttt tattaaaacg tctcaaaatc
      481 gtttctgaga cgttttagcg tttatttcgt ttagttatcg gcataatcgt taaaacaggc
      541 gttatcgtag cgtaaaagcc cttgagcgta gcgtgctttg cagcgaagat gttgtctgtt
      601 agattatgaa agccgatgac tgaatgaaat aataagcgca gcgtccttct atttcggttg
      661 gaggaggctc aagggagttt gagggaatga aattccctca tgggtttgat tttaaaaatt
      721 gcttgcaatt ttgccgagcg gtagcgctgg aaaatttttg aaaaaaattt ggaatttgga
      781 aaaaaatggg gggaaaggaa gcgaattttg cttccgtact acgacccccc attaagtgcc
      841 gagtgccaat ttttgtgcca aaaacgctct atcccaactg gctcaagggt ttgaggggtt
      901 tttcaatcgc caacgaatcg ccaacgtttt cgccaacgtt ttttataaat ctatatttaa
      961 gtagctttat tgttgttttt atgattacaa agtgatacac taattttata aaattatttg
     1021 attggagttt tttaaatggt gatttcagaa tcgaaaaaaa gagttatgat ttctctgaca
     1081 aaagagcaag ataaaaaatt aacagatatg gcgaaacaaa aaggtttttc aaaatctgcg
     1141 gttgcggcgt tagctataga agaatatgca agaaaggaat cagaacaaaa aaaataagcg
     1201 aaagctcgcg tttttagaag gatacgagtt ttcgctactt gtttttgata aggtaatata
     1261 tcatggctat taaaaatact aaagctagaa attttggatt tttattatat cctgactcaa
     1321 ttcctaatga ttggaaagaa aaattagaga gtttgggcgt atctatggct gtcagtcctt
     1381 tacacgatat ggacgaaaaa aaagataaag atacatggaa tagtagtgat gttatacgaa
     1441 atggaaagca ctataaaaaa ccacactatc acgttatata tattgcacga aatcctgtaa
     1501 caatagaaag cgttaggaac aagattaagc gaaaattggg gaatagttca gttgctcatg
     1561 ttgagatact tgattatatc aaaggttcat atgaatattt gactcatgaa tcaaaggacg
     1621 ctattgctaa gaataaacat atatacgaca aaaaagatat tttgaacatt aatgattttg
     1681 atattgaccg ctatataaca cttgatgaaa gccaaaaaag agaattgaag aatttacttt
     1741 tagatatagt ggatgactat aatttggtaa atacaaaaga tttaatggct tttattcgcc
     1801 ttaggggagc ggagtttgga attttaaata cgaatgatgt aaaagatatt gtttcaacaa
     1861 actctagcgc ctttagatta tggtttgagg gcaattatca gtgtggatat agagcaagtt
     1921 atgcaaaggt tcttgatgct gaaacggggg aaataaaatg acaaacaaag aaaaagagtt
     1981 atttgctgaa aatgaggaat taaaaaaaga aattaaggac ttaaaagagc gtattgaaag
     2041 atacagagaa atggaagttg aattaagtac aacaatctaa attcacttta gataaaaatt
     2101 taggaggcat atcaaatgaa ctttaataaa attgatttag acaattggaa gagaaaagag
     2161 atatttaatc attatttgaa ccaacaaacg acttttagta taaccacaga aattgatatt
     2221 agtgttttat accgaaacat aaaacaagaa ggatataaat tttaccctgc atttattttc
     2281 ttagtgacaa gggtgataaa ctcaaataca gcttttagaa ctggttacaa tagcgacgga
     2341 gagttaggtt attgggataa gttagagcca ctttatacaa tttttgatgg tgtatctaaa
     2401 acattctctg gtatttggac tcctgtaaag aatgacttca aagagtttta tgatttatac
     2461 ctttctgatg tagagaaata taatggttcg gggaaattgt ttcccaaaac acctatacct
     2521 gaaaatgctt tttctctttc tattattcct tggacttcat ttactgggtt taacttaaat
     2581 atcaataata atagtaatta ccttctaccc attattacag caggaaaatt cattaataaa
     2641 ggtaattcaa tatatttacc gctatcttta caggtacatc attctgtttg tgatggttat
     2701 catgcaggat tgtttatgaa ctctattcag gaattgtcag ataggcctaa tgactggctt
     2761 ttataatatg agataatgcc gactgtactt tttacagtcg gttttctaat gtcactaacc
     2821 tgccccgtta gttgaagaag gtttttatat tacagctcca agatctagtc ttataactat
     2881 actgacaagc gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat
     2941 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3001 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3061 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3121 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3181 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3241 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3301 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3361 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3421 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3481 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3541 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3601 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3661 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3721 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3781 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3841 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3961 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4021 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4081 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4141 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4201 actcttgttc caaacttgaa caacactcaa ccctatctcg ggctattctt ttgatttata
     4261 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4321 cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt
     4381 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4441 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4501 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4561 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4921 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4981 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5041 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5101 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtatt
//
