LOCUS       sequence-308811-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derived from pGPI-SceI; Trimethoprim resistant; carries I-SceI gene;
            contains 475 bp from both upstream and downstream flanking regions 
            between the FRT sites on pAHCTX1-rhadCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  AUTHORS   Hogan AM, Rahman ASMZ, Lightly TJ, Cardona ST
  TITLE     A broad-host-range CRISPRi toolkit for silencing gene expression in 
            Burkholderia.
  JOURNAL   ACS Synth Biol. 2019 Sep 6. doi: 10.1021/acssynbio.9b00232.
  PUBMED    31491085
REFERENCE   2  (bases 1 to 5431)
  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: "ACS Synth 
            Biol. 2019 Sep 6. doi: 10.1021/acssynbio.9b00232."
COMMENT     Sequence Label: pAH18
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            624..733
                     /note="incP origin of transfer"
     CDS             766..1137
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      1755..2143
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     complement(2604..2620)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2605..2621)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(2679..2726)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        join(3808..3813,3814..3830,3831..3836)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4162..4398
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     primer_bind     complement(5034..5053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5194..5216
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5355..5374
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggatcctttt tgtccggtgt tgggttgaag gtgaagccgg tcggggccgc agcgggggcc
       61 ggcttttcag ccttgccccc ctgcttcggc cgccgtggct ccggcgtctt gggtgccggc
      121 gcgggttccg cagccttggc ctgcggtgcg ggcacatcgg cgggcttggc cttgatgtgc
      181 cgcctggcgt gcgagcggaa cgtctcgtag gagaacttga ccttccccgt ttcccgcatg
      241 tgctcccaaa tggtgacgag cgcatagccg gacgctaacg ccgcctcgac atccgccctc
      301 accgccagga acgcaaccgc agcctcatca cgccggcgct tcttggccgc gcgggattca
      361 acccactcgg ccagctcgtc ggtgtagctc tttggcatcg tctctcgcct gtcccctcag
      421 ttcagtaatt tcctgcattt gcctgtttcc agtcggtaga tattccacaa aacagcaggg
      481 aagcagcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
      541 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
      601 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
      661 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
      721 cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc tgatgaaacc
      781 aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga cgaacgaaga
      841 gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct gctggccgtc
      901 ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga gctggcccgc
      961 atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac cgacgacccg
     1021 cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat cgaagagaag
     1081 caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga gccatgactt
     1141 ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc
     1201 catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc aaggcaagac
     1261 cgagcgcctg ggtcacgtgc gcgtcacgaa ctgcgaggca aacaccctgc ccgctgtcat
     1321 ggccgaggtg atggcgaccc agcacggcaa cacccgttcc gaggccgaca agacctatca
     1381 cctgctggtt agcttccgcg cgggagagaa gcccgacgcg gagacgttgc gcgcgattga
     1441 ggaccgcatc tgcgctgggc ttggcttcgc cgagcatcag cgcgtcagtg ccgtgcatca
     1501 cgacaccgac aacctgcaca tccatatcgc catcaacaag attcacccga cccgaaacac
     1561 catccatgag ccgtatcggg cctaccgcgc cctcgctgac ctctgcgcga cgctcgaacg
     1621 ggactacggg cttgagcgtg acaatcacga aacgcggcag cgcgtttccg agaaccgcgc
     1681 gaacgacatg gagcggcacg cgggcgtgga aagcctggtc ggctggatcc ggccacgatg
     1741 cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac gtactaagct
     1801 ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg tttaacgaac
     1861 taaaccctca tggctaacgt actaagctct catggctaac gtactaagct ctcatgtttc
     1921 acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac ttaaatagcc
     1981 tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa agcttttaag
     2041 gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc ttagagcctc
     2101 tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat tctgagttgc
     2161 tgttcgggca gttggtgatg ttgctgaatc gccatcgtta caccagtgat tcgttgccgc
     2221 caacatccag cgaaacgttg ctggataagc tgattacccg gctggcggct agcctgaaaa
     2281 gtccctttgc gctggataaa ttttgtgatg aggcatcgtg cagtgagcgc gttttgcgtc
     2341 agcaatttcg ccagcagact ggaatgacca tcaatcaata tctgcgacag gtcagagtgt
     2401 gtcatgcgca atatcttctc cagcatagcc gcctgttaat cagtgatatt tcgaccgaat
     2461 gtggctttga agatagtaac tatttttcgg tggtgtttac ccgggaaacc gggatgacgc
     2521 ccagccagtg gcgtcatctc aattcgcaga aagattaaat gcatatcgat gagctcgcac
     2581 gaacccagtt gacataagct tatcgatacc gtcgacctcg aggggggccg cgcgttggcc
     2641 gattcattaa tgcagcgaat tagcttcaaa agcgctctga agttcctata ctttctagag
     2701 aataggaact tcggaatagg aacttcaaga tcccctgatt ccctttgtca acagcaatgg
     2761 attttcaatg ggtcaacatg aaatccggct gttgccctgg gacggcaggt ttgacggcag
     2821 gcttccaggc tctctcgcac acgctgggaa cactctggga acaaacgcga gtgaaaatgg
     2881 acagtagggc ccaggaataa tgcggcctcc agcgaggttg gggcgtttca tatgagttcg
     2941 aatctcaccg cttccgccag gaagttcgcc gaaccccgtc cattcgcaga atgacggggt
     3001 ttttgtttgt ccgcgcgcgt gcgccccgca cttcccgacg cgccccgccg gatgccgtta
     3061 caatccggcg ctccgattcg cgaaacggga aatccacgtc atggtacctc tagaagaagc
     3121 ttgggatccg tcggggatcc ggtgattgat tgagcaagct ttatgcttgt aaaccgtttt
     3181 gtgaaaaaat ttttaaaata aaaaagggga cctctagggt ccccaattaa ttagtaatat
     3241 aatctattaa aggtcattca aaaggtcatc caccggatca gcttagtaaa gccctcgcta
     3301 gattttaatg cggatgttgc gattacttcg ccaactattg cgataacaag aaaaagccag
     3361 cctttcatga tatatctccc aatttgtgta gggcttatta tgcacgctta aaaataataa
     3421 aagcagactt gacctgatag tttggctgtg agcaattatg tgcttagtgc atctaacgct
     3481 tgagttaagc cgcgccgcga agcggcgtcg gcttgaacga attgttagac attatttgcc
     3541 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctagctt
     3601 taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct
     3661 aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg
     3721 gttatgccgg tactgccggg cctcttgcgg gattctacgg ggtctgacgc tcagtggaac
     3781 gaaatcgatg agctcgcacg aacccagttg acataagcct gttcggttcg taaactgtaa
     3841 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     3901 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca gtctatgcct
     3961 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     4021 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggcagccg ttgtgctggt
     4081 gctttctgat agttgttgtg gggtaggcag tcagagctcg atttgcttgt cgccataata
     4141 gattcacaag aaggattcga catgggtcaa agtagcgatg aagccaacgc tcccgttgca
     4201 gggcagtttg cgcttcccct gagtgccacc tttggcttag gggatcgcgt acgcaagaaa
     4261 tctggtgccg cttggcaggg tcaagtcgtc ggttggtatt gcacaaaact cactcctgaa
     4321 ggctatgcgg tcgagtccga atcccaccca ggctcagtgc aaatttatcc tgtggctgca
     4381 cttgaacgtg tggcctaagc attaccctgt tatccctaac tgcactcacc agtcacagaa
     4441 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4501 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4561 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4621 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     4681 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4741 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4801 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4861 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4921 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4981 tcagaccaag tttactcata tatactttag attgatttat ggtgcactct cagtacaatc
     5041 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     5101 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5161 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5221 caccgtcatc accgaaacgc gcgaggcagc aaggagatgg cgcccaacag tcccccggcc
     5281 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     5341 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     5401 gtgatgccgg ccacgatgcg tccggcgtag a
//
