LOCUS       sequence-296501-        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(650..661)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2914..3005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3006..3074,3075..3866)
                     /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(3224..3243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3908..4363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3995..4014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4205..4226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4474..5062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4963..4982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtattac cgcctttgag tgagcccgat gaagctgaca gctaaggtac cactagttct
       61 agagagctca agctttcttt gaaccaaaat tagaaaacca aggcttgaaa cgttcaattg
      121 aaatggcaat taaacaaatt acagcacgtg ttgctttgat tgatagccaa aaagcagcag
      181 ttgataaagc aattactgat attgctgaaa aattgtaatt tataaataaa aatcaccttt
      241 tagaggtggt ttttttattt ataaattatt cgtttgattt cgctttcgat agaacaatca
      301 aagcgagaat aaggaagata aatcccataa gggcgggagc agaatgtccg agactaatgt
      361 aaatttgtcc accaattaaa ggaccgataa cgcgagcttc tcgagtgcat attttcggca
      421 atcttctcaa tgagatgctc ttcagcatgt tcaatgatgt cgatttttta ttaaaacgtc
      481 tcaaaatcgt ttctgagacg ttttagcgtt tatttcgttt agttatcggc ataatcgtta
      541 aaacaggcgt tatcgtagcg taaaagccct tgagcgtagc gtggctttgc agcgaagatg
      601 ttgtctgtta gattatgaaa gccgatgact gaatgaaata ataagcgcag cgtccttcta
      661 tttcggttgg aggaggctca agggagtttg agggaatgaa attccctcat gggtttgatt
      721 ttaaaaattg cttgcaattt tgccgagcgg tagcgctgga aaatttttga aaaaaatttg
      781 gaatttggaa aaaaatgggg ggaaaggaag cgaattttgc ttccgtacta cgacccccca
      841 ttaagtgccg agtgccaatt tttgtgccaa aaacgctcta tcccaactgg ctcaagggtt
      901 tgaggggttt ttcaatcgcc aacgaatcgc caacgttttc gccaacgttt tttataaatc
      961 tatatttaag tagctttatt tttgttttta tgattacaaa gtgatacact aattttataa
     1021 aattatttga ttggagtttt ttaaatggtg atttcagaat cgaaaaaaag agttatgatt
     1081 tctctgacaa aagagcaaga taaaaaatta acagatatgg cgaaacaaaa agatttttca
     1141 aaatctgcgg ttgcggcgtt agctatagaa gaatatgcaa gaaaggaatc agaacaaaaa
     1201 aaataagcga aagctcgcgt ttttagaagg atacgagttt tcgctacttg tttttgataa
     1261 ggtaattata tcatggctat taaaaatact aaagttagaa attttggatt tttattatat
     1321 cctgactcaa ttcctaatga ttggaaagaa aaattagaga gtttgggcgt atctatggct
     1381 gtcagtcctt tacacgatat ggacgaaaaa aagataaaga tacatggaat agtagtgatg
     1441 ttatacgaaa tggaaagcac tataaaaaac cacactatca cgttatatat attgcacgaa
     1501 atcctgtaac aatagaaagc gttaggaaca agattaagcg aaaattgggg aatagttcag
     1561 ttgctcatgt tgagatactt gattatatca aaggttcata tgaatatttg actcatgaat
     1621 caaaggacgc tattgctaag aataaacata tatacgacaa aaaagatatt ttgaacatta
     1681 atgattttga tattgaccgc tatataacac ttgatgaaag ccaaaaaaga gaattgaaga
     1741 atttactttt agatatagtg gatgactata atttggtaaa tacaaaagat ttaatggctt
     1801 ttattcgcct taggggagcg gagtttggaa ttttaaatac gaatgatgta aaagatattg
     1861 tttcaacaaa ctctagcgcc tttagattat ggtttgaggg caattatcag tgtggatata
     1921 gagcaagtta tgcaaaggtt cttgatgctg aaacggggga aataaaatga caaacaaaga
     1981 aaaagagtta tttgctgaaa atgaggaatt aaaaaaagaa attaaggact taaaagagcg
     2041 tattgaaaga tacagagaaa tggaagttga attaagtaca acaatctaaa ttcactttag
     2101 ataaaaattt aggaggcata tcaaatgaac tttaataaaa ttgatttaga caattggaag
     2161 agaaaagaga tatttaatca ttatttgaac caacaaacga cttttagtat aaccacagaa
     2221 attgatatta gtgttttata ccgaaacata aaacaagaag gatataaatt ttaccctgca
     2281 tttattttct tagtgacaag ggtgataaac tcaaatacag cttttagaac tggttacaat
     2341 agcgacggag agttaggtta ttgggataag ttagagccac tttatacaat ttttgatggt
     2401 gtatctaaaa cattctctgg tatttggact cctgtaaaga atgacttcaa agagttttat
     2461 gatttatacc tttctgatgt agagaaatat aatggttcgg ggaaattgtt tcccaaaaca
     2521 cctatacctg aaaatgcttt ttctctttct attattcctt ggacttcatt tactgggttt
     2581 aacttaaata tcaataataa tagtaattac cttctaccca ttattacagc aggaaaattc
     2641 attaataaag gtaattcaat atatttaccg ctatctttac aggtacatca ttctgtttgt
     2701 gatggttatc atgctggatt gtttatgaac tctattcagg aattgtcaga taggcctaat
     2761 gactggcttt tataatatga gataatgccg actgtacttt ttacagtcgg ttttctaatg
     2821 tcactaacct gccccgttag ttgaagaagg tttttatatt acagctccag atctagtctt
     2881 ataactatac tgacaagcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     2941 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3001 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3061 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3121 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3181 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3241 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3301 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3361 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3421 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3481 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3541 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3601 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3661 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3721 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3781 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3841 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3901 ttgatttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3961 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4021 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4081 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4141 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4201 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     4261 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4321 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa
     4381 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4441 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4501 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4561 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4621 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4681 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4741 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4801 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4861 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4921 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4981 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5041 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5101 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggata
//
