LOCUS       sequence-426773-        7971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSCrhaB2plus with modifications of resistance cassette to CmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  AUTHORS   Yap ZL, Rahman ASMZ, Hogan AM, Levin DB, Cardona ST
  TITLE     A CRISPR-Cas-associated transposon system for genome editing in 
            Burkholderia cepacia complex species.
  JOURNAL   Appl Environ Microbiol. 2024 Jul 24;90(7):e0069924. doi: 
            10.1128/aem.00699-24. Epub 2024 Jun 13.
  PUBMED    38869300
REFERENCE   2  (bases 1 to 7971)
  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: "Appl 
            Environ Microbiol."; date: "2024-07-24"; volume: "90(7):e0069924. 
            doi"; pages: " 10.1128/aem.00699-24. Epub 2024 Jun 13"
COMMENT     Sequence Label: pZLrhaB2plus
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     341..360
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(1316..1418)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1535..1553)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      2811..3582
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3583..4245
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      complement(4711..4738)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4830..4916)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5069..5086
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             5483..6319
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     CDS             6393..7241
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     promoter        7398..7500
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
       61 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
      121 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gatatcggca
      181 ttcacacgct gaacgaaatc ttccaggttg agatcgatat cggggtactt tcacaaatta
      241 ttattctgag tgtttttaat tgtgatttcg ttttcttcat tgtgtttgtt gttaactctg
      301 tctctttaat ttttgtccag atatcttagg atctcccatc ggtgatgtcg gcgatatagg
      361 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
      421 tctgctcatg tttgacagct tatcatcgat ttcgttccac tgagcgtcag accccgtaga
      481 ccactcacta cagcagagcc atttaaacaa catcccctcc ccctttccac cgcgtcagac
      541 gcccgtagca gcccgctacg ggctttttca tgccctgccc tagcgtccaa gcctcacggc
      601 cgcgctcggc ctctctggcg gccttctggc gctgaggtct gcctcgtgaa gaaggtgttg
      661 ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg gagccacggt
      721 tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc tttgccacgg
      781 aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttcgat
      841 ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca
      901 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
      961 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1021 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1081 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1141 accatgagtg acgactgaat ccggtgagaa tggcaaaagc taaaaaggcc gtaatatcca
     1201 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     1261 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     1321 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     1381 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
     1441 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
     1501 cttccgtcac aggtatttat tcgaagacga aagggcctcg tgatacgcct atttttatag
     1561 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     1621 cgcgcccgcg ttcctgctgg cgctgggcct gtttctggcg ctggacttcc cgctgttccg
     1681 tcagcagctt ttcgcccacg gccttgatga tcgcggcggc cttggcctgc atatcccgat
     1741 tcaacggccc cagggcgtcc agaacgggct tcaggcgctc ccgaaggtct cgggccgtct
     1801 cttgggcttg atcggccttc ttgcgcatct cacgcgctcc tgcggcggcc tgtagggcag
     1861 gctcataccc ctgccgaacc gcttttgtca gccggtcggc cacggcttcc ggcgtctcaa
     1921 cgcgctttga gattcccagc ttttcggcca atccctgcgg tgcataggcg cgtggctcga
     1981 ccgcttgcgg gctgatggtg acgtggccca ctggtggccg ctccagggcc tcgtagaacg
     2041 cctgaatgcg cgtgtgacgt gccttgctgc cctcgatgcc ccgttgcagc cctagatcgg
     2101 ccacagcggc cgcaaacgtg gtctggtcgc gggtcatctg cgctttgttg ccgatgaact
     2161 ccttggccga cagcctgccg tcctgcgtca gcggcaccac gaacgcggtc atgtgcgggc
     2221 tggtttcgtc acggtggatg ctggccgtca cgatgcgatc cgccccgtac ttgtccgcca
     2281 gccacttgtg cgccttctcg aagaacgccg cctgctgttc ttggctggcc gacttccacc
     2341 attccgggct ggccgtcatg acgtactcga ccgccaacac agcgtccttg cgccgcttct
     2401 ctggcagcaa ctcgcgcagt cggcccatcg cttcatcggt gctgctggcc gcccagtgct
     2461 cgttctctgg cgtcctgctg gcgtcagcgt tgggcgtctc gcgctcgcgg taggcgtgct
     2521 tgagactggc cgccacgttg cccattttcg ccagcttctt gcatcgcatg atcgcgtatg
     2581 ccgccatgcc tgcccctccc ttttggtgtc caaccggctc gacgggggca gcgcaaggcg
     2641 gtgcctccgg cgggccactc aatgcttgag tatactcact agactttgct tcgcaaagtc
     2701 gtgaccgcct acggcggctg cggcgcccta cgggcttgct ctccgggctt cgccctgcgc
     2761 ggtcgctgcg ctcccttgcc agcccgtgga tatgtggacg atggccgcga gcggccaccg
     2821 gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg gacaggctgc
     2881 gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt cgaccacata
     2941 cccaccggct ccaactgcgc ggcctgcggc cttgccccat caattttttt aattttctct
     3001 ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg ccggttggac accaagtgga
     3061 aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg gccttgacgc gcctggaacg
     3121 acccaagcct atgcgagtgg gggcagtcga agggcgaagc ccgcccgcct gccccccgag
     3181 cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc caccttgggc aaggccgaag
     3241 gccgcgcagt cgatcaacaa gccccggagg ggccactttt tgccggaggg ggagccgcgc
     3301 cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg gcaccaaaga actagatata
     3361 gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa aggggggtac gcaacagctc
     3421 attgcggcac cccccgcaat agctcattgc gtaggttaaa gaaaatctgt aattgactgc
     3481 cacttttacg caacgcataa ttgttgtcgc gctgccgaaa agttgcagct gattgcgcat
     3541 ggtgccgcaa ccgtgcggca cccctaccgc atggagataa gcatggccac gcagtccaga
     3601 gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga acgcaaagcg
     3661 catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct gctgcatcac
     3721 ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac actttccaag
     3781 ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt ggccgagcgc
     3841 tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt ggtcaatgac
     3901 cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag tgccgccgtg
     3961 gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct gcgccgcatc
     4021 ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga gccgcccagc
     4081 cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac ggaggaatgg
     4141 gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct ggacgatggc
     4201 gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt gggttgcgca
     4261 gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg ccctatacct
     4321 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     4381 agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga ggcagaataa
     4441 atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg gcctcaggca
     4501 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     4561 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
     4621 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagt
     4681 cgaccaataa caagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     4741 ttaatttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     4801 ttcgcaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
     4861 aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     4921 ctggcagttc cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt
     4981 cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt
     5041 ctgttttatc agaccgcttc tgcgttctga tttaatctgt atcaggctga aaatcttctc
     5101 tcatccgcca aaacagccaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt
     5161 accatggcat atgtgatcct gctgaatttc actagaggga aaccgttgtg gtctccctta
     5221 cgaccagtct aaaaagcgcc tgaattcgcg acctttccct ggttgccagg aaaatgggcc
     5281 attggcaacc agggaaagat gaacgtgatg atgttcacaa tttgctgaat tgtggtgatg
     5341 tgatgctcac cgcatttcct gaaaattcac gctgtatctt gaaaaatcga cgttttttac
     5401 gtggttttcc gtcgaaaatt taaggtaaga acctgacctc gtgattacta tttcgccgtg
     5461 ttgacgacat caggaggcca gtatgaccgt attacatagt gtggattttt ttccgtctgg
     5521 taacgcgtcc gtggcgatag aaccccggct cccgcaggcg gattttcctg aacatcatca
     5581 tgattttcat gaaattgtga ttgtcgaaca tggcacgggt attcatgtgt ttaatgggca
     5641 gccctatacc atcaccggtg gcacggtctg tttcgtacgc gatcatgatc ggcatctgta
     5701 tgaacatacc gataatctgt gtctgaccaa tgtgctgtat cgctcgccgg atcgatttca
     5761 gtttctcgcc gggctgaatc agttgctgcc acaagagctg gatgggcagt atccgtctca
     5821 ctggcgcgtt aaccacagcg tattgcagca ggtgcgacag ctggttgcac agatggaaca
     5881 gcaggaaggg gaaaatgatt taccctcgac cgccagtcgc gagatcttgt ttatgcaatt
     5941 actgctcttg ctgcgtaaaa gcagtttgca ggagaacctg gaaaacagcg catcacgtct
     6001 caacttgctt ctggcctggc tggaggacca ttttgccgat gaggtgaatt gggatgccgt
     6061 ggcggatcaa ttttctcttt cactgcgtac gctacatcgg cagcttaagc agcaaacggg
     6121 actgacgcct cagcgatacc tgaaccgcct gcgactgatg aaagcccgac atctgctacg
     6181 ccacagcgag gccagcgtta ctgacatcgc ctatcgctgt ggattcagcg acagtaacca
     6241 cttttcgacg ctttttcgcc gagagtttaa ctggtcaccg cgtgatattc gccagggacg
     6301 ggatggcttt ctgcaataac gcgaatcttc tcaacgtatt tgtacgccat attgcgaata
     6361 atcaacttcg ttctctggcc gaggtagcca cggtggcgca tcagttaaaa cttctcaaag
     6421 atgatttttt tgccagcgac cagcaggcag tcgctgtggc tgaccgttat ccgcaagatg
     6481 tctttgctga acatacacat gatttttgtg agctggtgat tgtctggcgc ggtaatggcc
     6541 tgcatgtact caacgatcgc ccttatcgca ttacccgtgg cgatctcttt tacattcatg
     6601 ctgacgataa acactcctac gcttccgtta acgatctggt tttgcagaat attatttatt
     6661 gcccggagcg tctgaagctg aatcttgact ggcagggggc gattccggga tttaacgcca
     6721 gcgcagggca accacactgg cgcttaggta gcatggggat ggcgcaggcg cggcaggtta
     6781 tcggtcagct tgagcatgaa agtagtcagc atgtgccgtt tgctaacgaa atggctgagt
     6841 tgctgttcgg gcagttggtg atgttgctga atcgccatcg ttacaccagt gattcgttgc
     6901 cgccaacatc cagcgaaacg ttgctggata agctgattac ccggctggcg gctagcctga
     6961 aaagtccctt tgcgctggat aaattttgtg atgaggcatc gtgcagtgag cgcgttttgc
     7021 gtcagcaatt tcgccagcag actggaatga ccatcaatca atatctgcga caggtcagag
     7081 tgtgtcatgc gcaatatctt ctccagcata gccgcctgtt aatcagtgat atttcgaccg
     7141 aatgtggctt tgaagatagt aactattttt cggtggtgtt tacccgggaa accgggatga
     7201 cgcccagcca gtggcgtcat ctcaattcgc agaaagatta aatgcattct tctgacttct
     7261 tgtctgagaa aatgcataag tataggaact tcggcgcgcc tacctgtgac ggaagatcac
     7321 ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt
     7381 tggcgaaaat gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata
     7441 agatcactac cgggcgtatt ttttgagttg tcgagatttt caggagctaa ggaagctaaa
     7501 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     7561 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     7621 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     7681 cacattcttg cccgcctgat gaatgctcat ccggaattac gtatggcaat gaaagacggt
     7741 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     7801 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     7861 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     7921 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga t
//
