LOCUS       sequence-371382-       10679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VcDART under control of Lac promoter, Vc_2xBsaI_NT gRNA, 2xLguI 
            cargo stuffer, with ET-Seq (AsiSI+SbfI) restriction sites flanking 
            Tn7 Right End.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10679)
  AUTHORS   Rubin BE, Diamond S, Cress BF, Crits-Christoph A, Lou YC, Borges AL,
            Shivram H, He C, Xu M, Zhou Z, Smith SJ, Rovinsky R, Smock DCJ, Tang
            K, Owens TK, Krishnappa N, Sachdeva R, Barrangou R, Deutschbauer AM,
            Banfield JF, Doudna JA
  TITLE     Species- and site-specific genome editing in complex bacterial 
            communities.
  JOURNAL   Nat Microbiol. 2022 Jan;7(1):34-47. doi: 10.1038/s41564-021-01014-7.
            Epub 2021 Dec 6.
  PUBMED    34873292
REFERENCE   2  (bases 1 to 10679)
  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: 
            "10.1038/s41564-021-01014-7"; journalName: "Nat Microbiol"; date: 
            "2022-01"; volume: "7"; issue: "1"; pages: "34-47"
COMMENT     Sequence Label: pBFC0996
FEATURES             Location/Qualifiers
     source          1..10679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      310..698
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     promoter        join(848..853,854..871,872..878)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             879..890
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2975..2986
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     9481..9500
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9481..9499
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     9623..9643
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     oriT            9687..9795
                     /note="incP origin of transfer"
     promoter        9988..10092
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10093..10161,10162..10679,1..274)
                     /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(10311..10330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
       61 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      121 tgagcgtggc tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      181 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      241 tgagataggt gcctcactga ttaagcattg gtaactgtca gacccggcca cgatgcgtcc
      301 ggcgtagagg atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat
      361 gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac
      421 cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta
      481 ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa
      541 ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt ttaaggttta
      601 acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa
      661 gcaattttga gtgacacagg aacacttaac ggctgacatg ggcaagttta ctcatatata
      721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
      781 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgaca cctgcccgaa
      841 ccatatcttt acactttatg cttccggctc gtatgttgaa agaggagaaa ggatcaatgg
      901 cgacaagttt acctacgccc tcagcaatta cgacttcggc gttagagtat gcattccata
      961 ctcccgctcg caatctaacg aaatctcgcg gaaaaaacat tcatcgttat gtcagtgtaa
     1021 agatgagtaa gaggattacg gtagaatcta ctctagagtg tgatgcctgc tatcactttg
     1081 attttgagcc aagtattgtt cgcttttgcg ctcaaccgat tcgattttta tattatctca
     1141 atggtcagtc tcactcctat gttcctgact ttctagttca atttgatacc aacgagtttg
     1201 ttctatatga agtaaagtca gcttatgcta agaacaaacc tgattttgat gttgaatggg
     1261 aggcgaaagt aaaagcagca actgaactag ggctagaatt ggagcttgtt gaagagagtg
     1321 atattaggga tacggttgta ttaaataatc ttaagcgcat gcatcgttat gcttcgaaag
     1381 atgagctgaa taacgtacat aactctctct taaaaataat aaagtacaat ggcgcccaat
     1441 ctgcaagatg cttgggagaa cagttgggtt taaaaggccg aactgtttta ccaattttgt
     1501 gcgatttgct gtcaaggtgt ttactcgata cacgtttgga taagcctcta tctcttgaat
     1561 ctcgatttga gttggccagt tatggctaag aaagggttct caagtttcca tagaaaagca
     1621 gtctcttccc aagatacgct tgaatcgata gagcttgtct ctagcgctaa ttgtctagaa
     1681 agtgttacgt atcaagatat atcagcattt cccgaaacaa ttgcggtaga gattaatttc
     1741 cgattaagca ttcttcgctt tttagcgcga aagtgcgaaa ccattgtggc caaatcaatt
     1801 gaaccacatc gtgtagagct acagcaaaac tatagtagaa aaatacccag tgcaataacg
     1861 atatatcgat ggtggcttgc ttttcgaaaa tcagactaca accccattag cttagcacct
     1921 aatatcaagg atagaggtaa tagagagaca aaagtgtcaa cagttgttga ttctattatg
     1981 gaacaggcag ttgaaagagt tatatctgga cgaaaagtca atgttagctc tgcatataaa
     2041 cgtgttcgac gaaaagttcg tcaatacaat ctaactcatg gaacgaaata cacgtatcct
     2101 aagtacgaat ctgtaagaaa gcgagtaaaa aagaaaaccc catttgagtt attagccgca
     2161 gggaaagggg agagagtagc taagagagag tttcgccgaa tgggaaaaaa gatcctcacg
     2221 tctagcgtgc tagagagggt tgaaatagat cacactgtcg ttgacctttt tgcagtacac
     2281 gaagagtatc gaatcccatt gggccgacct tggcttactc aattggttga ttgttacagt
     2341 aaagctgtaa tcggttttta tttaggtttc gagcctccta gctatgtgtc ggtttccctt
     2401 gcacttaaga atgcaataca acgcaaagat gacttaatct cctcgtatga atcgatcgag
     2461 aatgaatggc tatgttatgg catcccagac ctactcgtaa ctgataatgg taaagagttt
     2521 ttgtcgaaag catttgatca agcatgtgaa tcactattga tcaatgtgca tcaaaataaa
     2581 gttgaaacgc ccgacaacaa acctcatgtt gaacgtaact acgggactat taatacttct
     2641 ctgttagacg atttacctgg gaaatccttc agccagtacc ttcaaagaga agggtacgac
     2701 tctgtgggag aagctaccct tacactcaat gagattagag aaatttactt aatttggttg
     2761 gtggatattt atcataaaaa acccaatcag agaggcacta attgtcctaa tgttgcgtgg
     2821 aaaaagggtt gtcaagaatg ggaaccagag gagttctctg gttctaaaga cgaattagac
     2881 tttaaatttg ctattgttga ttacaaacaa cttactaaag tagggataac tgtctacaaa
     2941 gaactgagtt atagcaatga ccgtttagct gaatatagag ggaagaaagg aaaccataaa
     3001 gttcagttca agtataaccc tgagtgtatg gcagttattt gggtgttgga tgaggatatg
     3061 aatgagtact ttacagttaa tgcgattgac tacgaatatg caagtagagt atcactttgg
     3121 caacataaat ataacatgaa atatcaagca gaactaaatt cagcagaata tgatgaggac
     3181 aaggaaattg atgcagaaat aaaaattgaa gaaatcgcag atcgttcaat tgttaagact
     3241 aacaaaatca gagctcggag gcgtggcgct aggcatcaag agaatagcgc aagggctaag
     3301 tcaatcagta atgcgaaccc ggcctcgata caaaaacatg aagatgaaat cgttagtgca
     3361 gataatgacg attgggatat tgattatgtc tgagaaatcg tcaaatgagt gaaacgcgtg
     3421 aggctcgtat atcaagagct aaaagggcat ttgtatccac accgagcgtt aggaaaatct
     3481 tgagttacat ggatagatgt agagatctat cagacctaga gtctgagcct acatgcatga
     3541 tggtctatgg tgcttcgggt gtaggtaaaa cgaccgtcat caagaaatac ttaaatcaaa
     3601 acagaagaga gtccgaagcc gggggcgata taataccggt tttgcatatt gagttgccag
     3661 acaatgcgaa gccagtagat gcagcaaggg aattgctggt tgaaatgggt gacccgctag
     3721 cactttatga aactgactta gctagattga cgaaaagact gactgaatta atccctgcgg
     3781 tcggcgtgaa gctgattatt atcgatgagt tccaacattt ggtggaagaa aggtcaaatc
     3841 gggttcttac ccaagtaggt aattggctaa aaatgatact taacaaaacg aaatgtccaa
     3901 ttgttatatt tggtatgcca tactcaaaag ttgtactgca agcaaactcg caacttcacg
     3961 ggcgattttc cattcaggtt gaactgcgcc cctttagcta ccagggaggt agaggtgtat
     4021 ttaaaacttt tttggaatac cttgataaag ccctaccttt tgaaaaacag gctggcttag
     4081 ccaacgaaag tttgcagaaa aaattgtatg cattctctca gggaaacatg cgttcgttga
     4141 gaaaccttat ttatcaagca tctatcgaag caattgataa tcagcatgaa acgataaccg
     4201 aagaagattt cgtttttgca tcgaagttga catcgggcga taaacccaac tcatggaaaa
     4261 atccttttga ggagggtgtt gaggtaacag aagatatgtt acgaccgcca ccaaaagata
     4321 ttggttggga ggactatttg agacattcaa ccccgagagt gagtaaacca ggtagaaata
     4381 aaaacttttt cgaataacct aggctgctgc caccgctgag caataaccat cttagtatat
     4441 tagttaagta taagaaggag atatacatat gtttttgcaa agacctaaac cttacagcga
     4501 tgaaagttta gaaagtttct ttatccgagt ggctaacaaa aatggctacg gtgatgtcca
     4561 tcgcttccta gaagccacta aacgattcct tcaagacatt gaccataatg gctatcaaac
     4621 ctttccgact gatataactc ggataaaccc atactcagct aaaaacagtt ccagcgcacg
     4681 aactgcgtca ttcctgaaac ttgcacaatt gacatttaat gaaccgccag agctacttgg
     4741 gttggcaatt aacagaacaa acatgaaata ctcgccgtca actagcgcgg ttgttagagg
     4801 tgcagaagtc tttcctcgca gtttactacg gacgcactcc atcccctgct gtcctttgtg
     4861 tctgcgagaa aatggctacg cctcctacct ttggcacttt caggggtacg aatactgcca
     4921 cagccataac gtacctttaa ttaccacttg tagctgtggt aaggagtttg actaccgagt
     4981 atctgggtta aagggcattt gctgcaaatg caaggagcct atcaccttaa ccagcaggga
     5041 gaacggtcat gaggcagcgt gtactgtttc aaactggctt gctggccatg aatctaaacc
     5101 tctgccaaat cttcctaaaa gctaccgatg gggtttagtt cattggtgga tgggtattaa
     5161 agatagcgag ttcgatcact tttcgttcgt tcaatttttc tcaaactggc caaggtcatt
     5221 ccactcgata atcgaagatg aagtagagtt caaccttgag catgctgttg tcagcacgtc
     5281 tgaattacga ctaaaagatc ttcttggtcg attgtttttc ggttcaattc ggttacctga
     5341 gcggaatctt caacacaata tcatccttgg tgagcttctc tgctatttag aaaatcgttt
     5401 atggcaagac aagggattaa tcgccaacct caaaatgaac gcgttagagg cgactgtaat
     5461 gttaaattgt agcctcgatc agattgcatc aatggttgaa caacgcatct tgaagccaaa
     5521 tcgaaaaagc aagcccaaca gccctcttga tgttaccgat tatctatttc atttcggcga
     5581 tattttctgt ctttggttag ctgagttcca aagcgatgag tttaaccgtt cgttttatgt
     5641 gtcgaggtgg taaatgcaaa ctctgaaaga actaatcgca tccaatcctg acgacttaac
     5701 aactgaactt aagagagcat ttcgtccact cacaccgcat attgcaattg atggtaatga
     5761 acttgacgca ctgacgatat tagtcaattt aaccgataag actgatgatc agaaagacct
     5821 gctcgatcga gccaaatgca agcaaaaact tcgagatgaa aaatggtggg ctagctgcat
     5881 aaattgcgtt aattacagac aaagccataa cccaaaattc ccggatatac gttctgaagg
     5941 cgtgattcga acccaagccc tgggtgaatt accgagtttc ctactctcgt cctcaaaaat
     6001 cccaccatac cattggtcat atagccatga ttcaaaatac gtcaacaaaa gcgcattcct
     6061 caccaatgag ttttgttggg atggtgagat ctcatgtttg ggtgagcttc ttaaagatgc
     6121 agatcaccca ctttggaata ctttgaaaaa gttaggttgt tctcaaaaaa cttgcaaagc
     6181 aatggcaaaa caactagctg atattactct cacgactatc aatgtcacac tcgcaccaaa
     6241 ttacctgact caaatatctc tccccgatag tgatacatcc tacatttcac tctctcctgt
     6301 tgcatcgcta tcgatgcaaa gccactttca tcagaggctc caagatgaaa atcggcatag
     6361 cgcgataacg cggtttagcc gaactaccaa catgggggtc actgcgatga catgtggcgg
     6421 cgcatttaga atgttaaagt ctggcgctaa gttttccagc ccccctcatc accgattaaa
     6481 cagtaaacga agttggttga cgtcagagca tgttcagtca ttaaaacagt accagcgcct
     6541 caacaaaagc ctcatacctg aaaactctcg gattgcactc cgtagaaaat acaaaatcga
     6601 gcttcaaaat atggtcagat cttggtttgc aatgcaagac catacccttg attcgaacat
     6661 acttatccaa catttgaatc atgacctatc ttacttagga gccacaaaac gttttgcata
     6721 cgatccagcg atgaccaagc tctttactga gcttttgaaa cgagagttat caaattcaat
     6781 caataatggt gagcaacaca ctaatggatc gtttttagtc ctaccgaata tcagggtttg
     6841 tggcgcaaca gctttaagct ccccggtaac ggtggggatt ccatcactta cagctttctt
     6901 tggcttcgtt cacgcatttg aacggaatat aaatcgcacc acctcatcgt ttcgtgttga
     6961 atcctttgcg atatgcgtcc atcaactaca tgtcgagaag cgaggtttga cagcagagtt
     7021 tgtggaaaaa ggcgacggga ctatatccgc tcccgcgacc cgggatgact ggcagtgtga
     7081 tgtcgtattt agccttattt tgaacaccaa ctttgctcaa catattgacc aagatacgtt
     7141 agttacatca ctaccaaagc gattggctcg gggttcagca aaaattgcga ttgatgactt
     7201 taaacatatc aactcattct cgacattaga aacagcgatc gaatctctgc caatagaagc
     7261 tggtaggtgg ttatcacttt acgcacagtc aaacaataat ctaagtgatc tattagcagc
     7321 catgacagag gaccatcagc tcatggcaag ctgcgtcggt taccacttgt tagaagaacc
     7381 caaagataaa ccaaactccc tcagaggtta caaacacgct atcgccgagt gcatcattgg
     7441 actcattaac tcaatcacct ttagctcaga gactgatccc aacacaatct tttggtcgct
     7501 aaagaactat caaaactacc tagtggtaca gccaaggagt atcaacgatg aaactaccga
     7561 caaatctagc ctatgagcgc tctatcgacc catcagatgt ctgttttttt gtcgtctggc
     7621 ccgatgatag aaaaacacct ttaacctaca attctcgtac tctgctcggg caaatggaag
     7681 cggcatcatt agcctatgat gtctcaggtc aaccaataaa aagtgccacc gctgaggcgt
     7741 tagctcaagg gaaccctcat caagttgatt tctgccacgt tccatacggt gcgagtcata
     7801 ttgaatgcag tttctccgtt tcgttttctt ctgaactacg tcaaccatat aagtgtaact
     7861 caagcaaagt taaacaaacg ctagtgcaat tagtcgagct ctacgaaacg aaaatcggct
     7921 ggactgagct agcaacccga tatttgatga acatttgcaa cggtaaatgg ctgtggaaaa
     7981 atacccgtaa agcctattgc tggaacattg tacttacacc ttggccttgg aacggggaaa
     8041 aggttggatt tgaagatatt cgtactaact acacctcacg gcaagacttt aaaaataata
     8101 aaaattggtc tgctatagtt gaaatgatca aaaccgcatt ttctagtact gatgggctgg
     8161 cgatatttga agtcagggcc accttgcact tgccaacgaa tgctatggtg cggccaagcc
     8221 aagttttcac agaaaaagaa agtggcagta aaagtaaatc taaaactcaa aacagtcgag
     8281 tttttcagag tacaactatt gatggtgaac gatcgccaat actaggggcc tttaaaacgg
     8341 gagcagctat tgcaaccatt gacgactggt atcctgaagc cactgagcca ctaagggtcg
     8401 gacggtttgg ggttcatcgc gaagatgtca cttgctaccg tcatccgtct accggaaaag
     8461 attttttctc gatattacaa caagcagagc actatattga agtgttgagc gccaacaaaa
     8521 ctcccgctca agaaactatc aacgacatgc actttttaat ggctaacctg attaagggtg
     8581 ggatgttcca gcataaagga gactgactgt gaaatggtat tataagacaa tcacctttct
     8641 gccagagttg tgcaacaacg agtcactggc tgcaaagtgt ctccgcgttc tgcatggatt
     8701 taactatcag tatgagacac gaaatatcgg cgtttcattt ccgctttggt gtgatgcaac
     8761 ggttggaaaa aagatttcat ttgtcagcaa gaacaagata gaactcgact tactacttaa
     8821 acaacactat ttcgtccaaa tggaacaact tcaatacttt catatatcca acactgttct
     8881 cgtcccagaa gattgtacat acgtttcctt tagacgctgt caatctatag ataagctcac
     8941 agcagcaggg ctggcaagga aaatcagacg cctggagaaa cgtgctctat ctagaggcga
     9001 gcaatttgac ccatcatctt ttgctcaaaa agagcatact gcaatagcgc actaccactc
     9061 acttggggag tccagcaaac agacgaaccg caactttcga ctcaatatca ggatgctctc
     9121 ggagcaaccc cgtgagggga actcgatttt tagtagctat ggcttatcaa attcagaaaa
     9181 ctcgtttcag cctgtaccct taatctgacc taggctgctg ccaccggtga actgccgagt
     9241 aggtagctga taacgagacc tcgtttacct atcggtctcg tgaactgccg agtaggtagc
     9301 tgataaccca aaccagatgt caacacagct actgttgatg caaccataaa gtgatattta
     9361 ataattattt ataatcagca acttaaccac aaaacaacca tatattgata tctcacaaaa
     9421 caaccataag ttgatatttt tcagaaagga agagcgagat ctcgagctct tcggctgctg
     9481 taatacgact cactataggg gataagagac ggctcaacgc ccgtctcaca gcgcaatatc
     9541 aatttatggg tgtgataatt atcaatttat gggtgtaatt atcattttat ggttgtatca
     9601 acaaggagga ctttggcagg tgcagcacaa aaggaaactc accgcgatcg ccattactcg
     9661 catcagcctg caggattctt gcttatcggc cagcctcgca gagcaggatt cccgttgagc
     9721 accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact
     9781 tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt
     9841 tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg
     9901 accccgaagc agggttatgc agcggaaaag aatgtcatga taataatggt ttcttagacg
     9961 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
    10021 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
    10081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
    10141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
    10201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
    10261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
    10321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
    10381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
    10441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
    10501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
    10561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
    10621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaact
//
