LOCUS       Exported                6499 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Derived from pDAI-SceI. Tetracycline resistance removed and replaced
            with chloramphenicol resistance cassette from pKD3..
ACCESSION   .
VERSION     .
KEYWORDS    pAH25-SceI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6499)
  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 6499)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6499
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(137..165)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        303..405
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(1351..2013)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQRRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2014..2785
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     primer_bind     4043..4061
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4178..4280
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5134..5161
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5253..5339
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5492..5509
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(5567..5581)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc
       61 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
      121 acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcgag
      181 ctcatcgcta gcatactcga gcgcctacct gtgacggaag atcacttcgc agaataaata
      241 aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac
      301 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
      361 gtattttttg agttgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
      421 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
      481 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
      541 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
      601 ctgatgaatg ctcatccgga attacgtatg gcaatgaaag acggtgagct ggtgatatgg
      661 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
      721 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
      781 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
      841 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
      901 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
      961 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     1021 aatgaattac aacagtactg cgatgagtac cggtttattg actaccggaa gcagtgtgac
     1081 cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggtaataat
     1141 tgacgatatg atcatttatt ctgcctccca gagcctgata aaaacggtga atccgttagc
     1201 gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac
     1261 gcggggaggc agacaaggta tagggcggcg aggcggctac agccgatagt ctggaacagc
     1321 gcacttacgg gttgctgcgc aacccaagtg ctaccggcgc ggcagcgtga cccgtgtcgg
     1381 cggctccaac ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgccg
     1441 ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc
     1501 cggaatgccg ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc
     1561 cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc
     1621 gtcgtgatca accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg
     1681 gccccacgcc acgcggtcat tgaccacgta ggccgacacg gtgccggggc cgttgagctt
     1741 cacgacggag atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa
     1801 agaacgtccg atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc
     1861 catctgcgcc acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc
     1921 ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc
     1981 ttgaatgccg atttctctgg actgcgtggc catgcttatc tccatgcggt aggggtgccg
     2041 cacggttgcg gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg
     2101 cgttgcgtaa aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc
     2161 ggggggtgcc gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc
     2221 tttcgcattt cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt
     2281 tcccccacgc cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt
     2341 gatcgactgc gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca
     2401 ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac tgcccccact
     2461 cgcataggct tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg
     2521 agccttgacc cgccttccac ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc
     2581 cggaggcttt tccccagaga aaattaaaaa aattgatggg gcaaggccgc aggccgcgca
     2641 gttggagccg gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc cctgtggtca
     2701 agctcgtggg caggcgcagc ctgtccatca gcttgtccag cagggttgtc cacgggccga
     2761 gcgaagcgag ccagccggtg gccgctcgcg gccatcgtcc acatatccac gggctggcaa
     2821 gggagcgcag cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg cgccgcagcc
     2881 gccgtaggcg gtcacgactt tgcgaagcaa agtctagtga gtatactcaa gcattgagtg
     2941 gcccgccgga ggcaccgcct tgcgctgccc ccgtcgagcc ggttggacac caaaagggag
     3001 gggcaggcat ggcggcatac gcgatcatgc gatgcaagaa gctggcgaaa atgggcaacg
     3061 tggcggccag tctcaagcac gcctaccgcg agcgcgagac gcccaacgct gacgccagca
     3121 ggacgccaga gaacgagcac tgggcggcca gcagcaccga tgaagcgatg ggccgactgc
     3181 gcgagttgct gccagagaag cggcgcaagg acgctgtgtt ggcggtcgag tacgtcatga
     3241 cggccagccc ggaatggtgg aagtcggcca gccaagaaca gcaggcggcg ttcttcgaga
     3301 aggcgcacaa gtggctggcg gacaagtacg gggcggatcg catcgtgacg gccagcatcc
     3361 accgtgacga aaccagcccg cacatgaccg cgttcgtggt gccgctgacg caggacggca
     3421 ggctgtcggc caaggagttc atcggcaaca aagcgcagat gacccgcgac cagaccacgt
     3481 ttgcggccgc tgtggccgat ctagggctgc aacggggcat cgagggcagc aaggcacgtc
     3541 acacgcgcat tcaggcgttc tacgaggccc tggagcggcc accagtgggc cacgtcacca
     3601 tcagcccgca agcggtcgag ccacgcgcct atgcaccgca gggattggcc gaaaagctgg
     3661 gaatctcaaa gcgcgttgag acgccggaag ccgtggccga ccggctgaca aaagcggttc
     3721 ggcaggggta tgagcctgcc ctacaggccg ccgcaggagc gcgtgagatg cgcaagaagg
     3781 ccgatcaagc ccaagagacg gcccgagacc ttcgggagcg cctgaagccc gttctggacg
     3841 ccctggggcc gttgaatcgg gatatgcagg ccaaggccgc cgcgatcatc aaggccgtgg
     3901 gcgaaaagct gctgacggaa cagcgggaag tccagcgcca gaaacaggcc cagcgccagc
     3961 aggaacgcgg gcgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4021 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcgaataaa
     4081 tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg
     4141 ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta agaggttcca
     4201 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     4261 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     4321 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     4381 accagaccgt tcagctggat attacggcct ttttagcttt tgccattctc accggattca
     4441 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     4501 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     4561 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatacggt
     4621 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4681 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     4741 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     4801 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     4861 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     4921 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgccag aaggccgcca
     4981 gagaggccga gcgcggccgt gaggcttgga cgctagggca gggcatgaaa aagcccgtag
     5041 cgggctgcta cgggcgtctg acgcggtgga aagggggagg ggatgttgtt taaatggctc
     5101 tgtgtagtga gtggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     5161 tgcttaattt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     5221 tgcttcgcag cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     5281 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
     5341 tgcctggcag ttccctactc tcgcatgggg agaccccaca ctaccatcgg cgctacggcg
     5401 tttcacttct gagttcggca tggggtcagg tgggaccacc gcgctactgc cgccaggcaa
     5461 attctgtttt atcagaccgc ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt
     5521 ctctcatccg ccaaaacagc caagcttgca tgcctgcagg tcgtcacttg tcgtcgtcgt
     5581 cgtccttgta gtctctagat tatttcagga aagtttcgga ggagatagtg ttcggcagtt
     5641 tgtacatcat ctgcgggatc aggtacggtt tgatcaggtt gtagaagatc aggtaagaca
     5701 tagaatcgat gtagatgatc ggtttgtttt tgttgatttt tacgtaacag ttcagttgga
     5761 atttgttacg cagaccctta accaggtatt ctacttcttc gaaagtgaaa gactgggtgt
     5821 tcagtacgat cgatttgttg gtagagtttt tgttgtaatc ccatttacca ccatcatcca
     5881 tgaaccagta tgccagagac atcggggtca ggtagttttc aaccaggttg ttcgggatgg
     5941 tttttttgtt gttaacgatg aacaggttag ccagtttgtt gaaagcttgg tgtttgaaag
     6001 tctgggcgcc ccaggtgatt accaggttac ccaggtggtt aacacgttct tttttgtgcg
     6061 gcggggacag tacccactga tcgtacagca gacatacgtg gtccatgtat gctttgtttt
     6121 tccactcgaa ctgcatacag taggttttac cttcatcacg agaacggatg taagcatcac
     6181 ccaggatcag accgatacct gcttcgaact gttcgatgtt cagttcgatc agctgggatt
     6241 tgtattcttt cagcagttta gagttcggac ccaggttcat tacctggttt ttttgatgca
     6301 tatgtagaat tccgaatcct tcttgtgaat ctattatggc gacaagcaaa tcgagctctg
     6361 actgcctacc ccacaacaac tatcagaaag caccagcaca acggctgcct aactttgttt
     6421 tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca
     6481 cggcgtaacg cgcttgctg
//
