LOCUS       Exported                6496 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 6496)
  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 6496)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6496
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        36..64
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1119..1133
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(1191..1208)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1361..1447
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1539..1566
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        complement(2421..2523)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2640..2658)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3916..4685
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             4686..5348
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     promoter        complement(6292..6394)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tcgagtatgc tagcgatgag ctcgcacgaa cccagttgac ataagcctgt tcggttcgta
       61 aactgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
      121 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttgtacagt
      181 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
      241 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggcagccgtt
      301 gtgctggtgc tttctgatag ttgttgtggg gtaggcagtc agagctcgat ttgcttgtcg
      361 ccataataga ttcacaagaa ggattcggaa ttctacatat gcatcaaaaa aaccaggtaa
      421 tgaacctggg tccgaactct aaactgctga aagaatacaa atcccagctg atcgaactga
      481 acatcgaaca gttcgaagca ggtatcggtc tgatcctggg tgatgcttac atccgttctc
      541 gtgatgaagg taaaacctac tgtatgcagt tcgagtggaa aaacaaagca tacatggacc
      601 acgtatgtct gctgtacgat cagtgggtac tgtccccgcc gcacaaaaaa gaacgtgtta
      661 accacctggg taacctggta atcacctggg gcgcccagac tttcaaacac caagctttca
      721 acaaactggc taacctgttc atcgttaaca acaaaaaaac catcccgaac aacctggttg
      781 aaaactacct gaccccgatg tctctggcat actggttcat ggatgatggt ggtaaatggg
      841 attacaacaa aaactctacc aacaaatcga tcgtactgaa cacccagtct ttcactttcg
      901 aagaagtaga atacctggtt aagggtctgc gtaacaaatt ccaactgaac tgttacgtaa
      961 aaatcaacaa aaacaaaccg atcatctaca tcgattctat gtcttacctg atcttctaca
     1021 acctgatcaa accgtacctg atcccgcaga tgatgtacaa actgccgaac actatctcct
     1081 ccgaaacttt cctgaaataa tctagagact acaaggacga cgacgacgac aagtgacgac
     1141 ctgcaggcat gcaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag
     1201 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     1261 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     1321 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     1381 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1441 gacaaatccg ccgggagcgg atttgaacgc tgcgaagcaa cggcccggag ggtggcgggc
     1501 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     1561 cctttttgcg tttctacaaa ctcttccact cactacagca gagccattta aacaacatcc
     1621 cctccccctt tccaccgcgt cagacgcccg tagcagcccg ctacgggctt tttcatgccc
     1681 tgccctagcg tccaagcctc acggccgcgc tcggcctctc tggcggcctt ctggcgctga
     1741 ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc
     1801 agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg
     1861 attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga
     1921 tccttcaact cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt caagtcagcg
     1981 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     2041 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     2101 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2161 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2221 aaataaggtt atcaagcgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     2281 aaagctaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     2341 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     2401 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     2461 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     2521 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     2581 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcgaa gacgaaaggg
     2641 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2701 aggtggcact tttcggggaa atgtgcgcgc ccgcgttcct gctggcgctg ggcctgtttc
     2761 tggcgctgga cttcccgctg ttccgtcagc agcttttcgc ccacggcctt gatgatcgcg
     2821 gcggccttgg cctgcatatc ccgattcaac ggccccaggg cgtccagaac gggcttcagg
     2881 cgctcccgaa ggtctcgggc cgtctcttgg gcttgatcgg ccttcttgcg catctcacgc
     2941 gctcctgcgg cggcctgtag ggcaggctca tacccctgcc gaaccgcttt tgtcagccgg
     3001 tcggccacgg cttccggcgt ctcaacgcgc tttgagattc ccagcttttc ggccaatccc
     3061 tgcggtgcat aggcgcgtgg ctcgaccgct tgcgggctga tggtgacgtg gcccactggt
     3121 ggccgctcca gggcctcgta gaacgcctga atgcgcgtgt gacgtgcctt gctgccctcg
     3181 atgccccgtt gcagccctag atcggccaca gcggccgcaa acgtggtctg gtcgcgggtc
     3241 atctgcgctt tgttgccgat gaactccttg gccgacagcc tgccgtcctg cgtcagcggc
     3301 accacgaacg cggtcatgtg cgggctggtt tcgtcacggt ggatgctggc cgtcacgatg
     3361 cgatccgccc cgtacttgtc cgccagccac ttgtgcgcct tctcgaagaa cgccgcctgc
     3421 tgttcttggc tggccgactt ccaccattcc gggctggccg tcatgacgta ctcgaccgcc
     3481 aacacagcgt ccttgcgccg cttctctggc agcaactcgc gcagtcggcc catcgcttca
     3541 tcggtgctgc tggccgccca gtgctcgttc tctggcgtcc tgctggcgtc agcgttgggc
     3601 gtctcgcgct cgcggtaggc gtgcttgaga ctggccgcca cgttgcccat tttcgccagc
     3661 ttcttgcatc gcatgatcgc gtatgccgcc atgcctgccc ctcccttttg gtgtccaacc
     3721 ggctcgacgg gggcagcgca aggcggtgcc tccggcgggc cactcaatgc ttgagtatac
     3781 tcactagact ttgcttcgca aagtcgtgac cgcctacggc ggctgcggcg ccctacgggc
     3841 ttgctctccg ggcttcgccc tgcgcggtcg ctgcgctccc ttgccagccc gtggatatgt
     3901 ggacgatggc cgcgagcggc caccggctgg ctcgcttcgc tcggcccgtg gacaaccctg
     3961 ctggacaagc tgatggacag gctgcgcctg cccacgagct tgaccacagg gattgcccac
     4021 cggctaccca gccttcgacc acatacccac cggctccaac tgcgcggcct gcggccttgc
     4081 cccatcaatt tttttaattt tctctgggga aaagcctccg gcctgcggcc tgcgcgcttc
     4141 gcttgccggt tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga ccgcgcagcg
     4201 gcttggcctt gacgcgcctg gaacgaccca agcctatgcg agtgggggca gtcgaaggcg
     4261 aagcccgccc gcctgccccc cgagcctcac ggcggcgagt gcgggggttc caagggggca
     4321 gcgccacctt gggcaaggcc gaaggccgcg cagtcgatca acaagccccg gaggggccac
     4381 tttttgccgg agggggagcc gcgccgaagg cgtgggggaa ccccgcaggg gtgcccttct
     4441 ttgggcacca aagaactaga tatagggcga aatgcgaaag acttaaaaat caacaactta
     4501 aaaaaggggg gtacgcaaca gctcattgcg gcaccccccg caatagctca ttgcgtaggt
     4561 taaagaaaat ctgtaattga ctgccacttt tacgcaacgc ataattgttg tcgcgctgcc
     4621 gaaaagttgc agctgattgc gcatggtgcc gcaaccgtgc ggcaccctac cgcatggaga
     4681 taagcatggc cacgcagtcc agagaaatcg gcattcaagc caagaacaag cccggtcact
     4741 gggtgcaaac ggaacgcaaa gcgcatgagg cgtgggccgg gcttattgcg aggaaaccca
     4801 cggcggcaat gctgctgcat cacctcgtgg cgcagatggg ccaccagaac gccgtggtgg
     4861 tcagccagaa gacactttcc aagctcatcg gacgttcttt gcggacggtc caatacgcag
     4921 tcaaggactt ggtggccgag cgctggatct ccgtcgtgaa gctcaacggc cccggcaccg
     4981 tgtcggccta cgtggtcaat gaccgcgtgg cgtggggcca gccccgcgac cagttgcgcc
     5041 tgtcggtgtt cagtgccgcc gtggtggttg atcacgacga ccaggacgaa tcgctgttgg
     5101 ggcatggcga cctgcgccgc atcccgaccc tgtatccggg cgagcagcaa ctaccgaccg
     5161 gccccggcga ggagccgccc agccagcccg gcattccggg catggaacca gacctgccag
     5221 ccttgaccga aacggaggaa tgggaacggc gcgggcagca gcgcctgccg atgcccgatg
     5281 agccgtgttt tctggacgat ggcgagccgt tggagccgcc gacacgggtc acgctgccgc
     5341 gccggtagca cttgggttgc gcagcaaccc gtaagtgcgc tgttccagac tatcggctgt
     5401 agccgcctcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     5461 cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc accgttttta
     5521 tcaggctctg ggaggcagaa taaatgatca tatcgtcaat tattacctcc acggggagag
     5581 cctgagcaaa ctggcctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     5641 ataaaccggt actcatcgca gtactgttgt attcattaag catctgccga catggaagcc
     5701 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     5761 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     5821 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     5881 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     5941 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     6001 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     6061 cattgccata cgtaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     6121 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     6181 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     6241 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     6301 cttagctcct gaaaatctcg acaactcaaa aaatacgccc ggtagtgatc ttatttcatt
     6361 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc
     6421 ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc
     6481 cgtcacaggt aggcgc
//
