LOCUS       sequence-369168-        4530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KanR-BBR1 entry vector for HR donor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4530)
  AUTHORS   Adler BA, Hessler T, Cress BF, Lahiri A, Mutalik VK, Barrangou R, 
            Banfield J, Doudna JA
  TITLE     Broad-spectrum CRISPR-Cas13a enables efficient phage genome editing.
  JOURNAL   Nat Microbiol. 2022 Dec;7(12):1967-1979. doi: 
            10.1038/s41564-022-01258-x. Epub 2022 Oct 31.
  PUBMED    36316451
REFERENCE   2  (bases 1 to 4530)
  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/s41564-022-01258-x"; journalName: "Nat Microbiol"; date: 
            "2022-12"; volume: "7"; issue: "12"; pages: "1967-1979"
COMMENT     Sequence Label: pBA707
FEATURES             Location/Qualifiers
     source          1..4530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      309..380
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      396..423
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(484..1299)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(484..1299)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1207..1226
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1291..1310
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3323..3985
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     protein_bind    4065..4083
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4090..4108
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4126..4137
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(4144..4530,1..291)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt
       61 atgtacccgg aggacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac
      121 ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg
      181 ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga ggactacacc
      241 atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataaggatct
      301 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
      361 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
      421 gcgtttatag aagaccaagg aggactttgg caggtggtgt tacaaccaat taaccaattc
      481 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
      541 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      601 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
      661 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      721 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
      781 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
      841 ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
      901 cgaatgcaac cggcgcagga acactgcaag cgcatcaaca atattttcac ctgaatcagg
      961 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     1021 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     1081 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     1141 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     1201 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     1261 cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     1321 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     1381 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     1441 ggatcagctt gcgttttccc ttgtccagat agcccagtag ctgacattta gcctaaatga
     1501 catttctccg gaaagtgcgc acggccacgc tcttgttgac gttgagcttg cttttgacgt
     1561 tgaacctcgc gttgctcggt taataatttc tcaccaaccg ctttaataat ggccgccgct
     1621 ttcgcttgca tgtcacggtt cagaggaccg agtgcatcta ataccggttt caaacgttcg
     1681 cgtaaatcgc gcgcagtttc ctgtgcctgg tctgctttct tacgcatttc gcgggcgccg
     1741 gcagccgctt gcagtgctgg ttcgtaacct tgacggactg ccttagttaa gcgatccgcg
     1801 actgcctcag gggtttccac acgtttagaa atgcctaatt tctccgccag accttgtgga
     1861 gcgtaagcac ggggttccac ggcctgaggc gaaatcgtaa catgtcctac aggcgggcgt
     1921 tcaagcgctt cataaaatgc ttggatacga gtatggcgcg ctttactccc ttcaatccca
     1981 cgctgaaggc ccaggtccgc aaccgcagca gcgaaggtcg tttgatcacg tgtcatttgg
     2041 gccttattac caataaattc tttagcactc aagcgaccat cttgggtcaa ggggactaca
     2101 aaggccgtca tatgtgggga tgtctcatcg cgatgaatcg aagctgttac aatacggtca
     2161 gcgccatatt tatcggctaa ccatttatgg gctttctcaa agaatgctgc ttgttgctcc
     2221 tgtgacgcgc ttttccacca ctcgggcgaa gcggtcatca catattctac ggcaagtact
     2281 gcatctttac ggcgcttttc aggcaacagt tcacgcaaac gacccatggc ctcgtccgta
     2341 ctactcgcgg cccaatgttc attttccgga gtacgagatg catccgcatt cggtgtttca
     2401 cgttcacgat acgcatgttt cagcgacgca gcaacattac ccatcttggc gagtttctta
     2461 caacgcataa ttgcataagc ggccatgcct gcccctccct tttggtgtcc aaccggctcg
     2521 acgggggcag cgcaaggcgg tgcctccggc gggccactca atgcttgagt atactcacta
     2581 gactttgctt cgcaaagtcg tgaccgccta cggcggctgc gcctgcccac gagcttgacc
     2641 acagggattg cccaccggct acccagcctt cgaccacata cccaccggct ccaactgcgc
     2701 ggcctgcggc cttgccccat caattttttt aattttctct ggggaaaagc ctccggcctg
     2761 cggcctgcgc gcttcgcttg ccggttggac accaagtgga aggcgggtca aggctcgcgc
     2821 agcgaccgcg cagcggcttg gccttgacgc gcctggaacg acccaagcct atgcgagtgg
     2881 gggcagtcga agggcgaagc ccgcccgcct gccccccgag cctcacggcg gcgagtgcgg
     2941 gggttccaag ggggcagcgc caccttgggc aaggccgaag gccgcgcagt cgatcaacaa
     3001 gccccggagg ggccactttt tgccggaggg ggagccgcgc cgaaggcgtg ggggaacccc
     3061 gcaggggtgc ccttctttgg gcaccaaaga actagatata gggcgaaatg cgaaagactt
     3121 aaaaatcaac aacttaaaaa aggggggtac gcaacagctc attgcggcac cccccgcaat
     3181 agctcattgc gtaggttaaa gaaaatctgt aattgactgc cacttttacg caacgcataa
     3241 ttgttgtcgc gctgccgaaa agttgcagct gattgcgcat ggtgccgcaa ccgtgcggca
     3301 cccctaccgc atggagataa gcatggccac gcagtccaga gaaatcggca ttcaagccaa
     3361 gaacaagccc ggtcactggg tgcaaacgga acgcaaagcg catgaggcgt gggccgggct
     3421 tattgcgagg aaacccacgg cggcaatgct gctgcatcac ctcgtggcgc agatgggcca
     3481 ccagaacgcc gtggtggtca gccaaaagac actttccaag ctcatcggac gttctttgcg
     3541 gacggtccaa tacgcagtca aggacttggt ggccgagcgc tggatctccg tcgtgaagct
     3601 caacggcccc ggcaccgtgt tggcctacgt ggtcaatgac cgcgtggcgt ggggccagcc
     3661 ccgcgaccag ttgcgcctgt cggtgttcag tgccgccgtg gtggttgatc acgacgacca
     3721 ggacgaatcg ctgttggggc atggcgacct gcgccgcatc ccgaccctgt atccgggcga
     3781 gcagcaacta ccgaccggcc ccggcgagga gccgcccagc cagcccggca ttccgggcat
     3841 ggaaccagac ctgccagcct tgaccgaaac ggaggaatgg gaacggcgcg ggcagcagcg
     3901 cctgccgatg cccgatgagc cgtgttttct ggacgatggc gagccgttgg agccgccgac
     3961 acgggtcacg ctgccgcgcc ggtagcactt gggttgcgca gcaacccgta agtgcgctgt
     4021 tccagactat ccacctgccc gaaccagaga cttatcatgt cttctcccta tcagtgatag
     4081 agattgacat ccctatcagt gatagagata ctgagcacgg atctgaaaga ggagaaatga
     4141 tctatggcga gtagcgaaga tgttatcaaa gagttcatgc gtttcaaagt tcgtatggaa
     4201 ggttccgtta acggtcacga gttcgaaatc gaaggtgaag gtgaaggtcg tccgtacgaa
     4261 ggtactcaga ccgctaaact gaaagttacc aaaggtggtc cgctgccgtt cgcttgggac
     4321 atcctgtccc cgcagttcca gtacggttcc aaagcttacg ttaaacaccc ggctgacatc
     4381 ccggactacc tgaaactgtc cttcccggaa ggtttcaaat gggaacgtgt tatgaacttc
     4441 gaggacggtg gtgttgttac cgttacccag gactcctccc tgcaagacgg tgagttcatc
     4501 tacaaagtta aactgcgtgg tactaacttc
//
