LOCUS       sequence-301659-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with a mutated IHFprox site, that 
            should not be recognized by IHF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  AUTHORS   Santiago-Frangos A, Buyukyoruk M, Wiegand T, Krishna P, Wiedenheft B
  TITLE     Distribution and phasing of sequence motifs that facilitate CRISPR 
            adaptation.
  JOURNAL   Curr Biol. 2021 Aug 23;31(16):3515-3524.e6. doi: 
            10.1016/j.cub.2021.05.068. Epub 2021 Jun 25.
  PUBMED    34174210
REFERENCE   2  (bases 1 to 7044)
  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: 
            "10.1016/j.cub.2021.05.068"; journalName: "Curr Biol"; date: 
            "2021-08-23- 23"; volume: "31"; issue: "16"; pages: "3515-3524.e6"
COMMENT     Sequence Label: pCRISPR2_IHF_Mut
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        670..954
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     877..896
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             981..1003
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2938..2954)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2947..2969)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3115..3948)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      3962..4313
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     promoter        join(4354..4359,4360..4376,4377..4382)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4552..5085
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      5886..6474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6375..6394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            6544..6652
                     /note="incP origin of transfer"
     CDS             complement(join(6809..7044,1..643))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
       61 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
      121 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
      181 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
      241 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
      301 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
      361 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
      421 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
      481 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
      541 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa
      601 acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc
      661 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
      721 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
      781 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
      841 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
      901 gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt
      961 ttttttgggc tagaaataat tttgtttaac tttaagaagg agatatacat acccatggat
     1021 gccttgtccg aaatcgaggc gggccacccg agcctgtagc gccgctccct ggcgcgacgc
     1081 ccccggcctg gccgggggct ttgcggcatc gcccatcaaa agacctttcg cgcccgaacg
     1141 gcacggttga tcgccgtccc ggtcctcgcg aaacggccgc caattgcccg aagcttccga
     1201 cccttttttc ggacgatttc ttacgccctt ataaatcagc aagttacgag acctcgaaaa
     1261 aagagggttt ctggcgggaa aaactcggta tttctccttc cttcccatgg atttaggttt
     1321 tcggagctag ttcactgccg tgtaggcagc taagaaaatc agccggacgt tgtagtagtc
     1381 gagcgcggtg ttcactgccg tgtaggcagc taagaaagcc gggtggtgac ttctggacgg
     1441 cgttcgtcaa gttcactgcc gtgtaggcag ctaagaaaga cctggttttg gccgacacag
     1501 tggctgcgac gttcactgcc gtgtaggcag ctaagaaacg gtcctgccag tactgcggca
     1561 ggatctggtc gttcactgcc gtgtaggcag ctaagaaaag cctggcgccc ttcgagtcca
     1621 tccagcccgg gttcactgcc gtgtaggcag ctaagaaaca tccaattcgc cgatccggcg
     1681 caagcgcccg gttcactgcc gtgtaggcag ctaagaaaac gaagcagatc gccgaatgcg
     1741 gcgtcagcgc gttcactgcc gtgtaggcag ctaagaaagt gggtgcaaat cgcgattctg
     1801 gtcgcgtcgt gttcactgcc gtgtaggcag ctaagaaatg aatatcagcc tgcaaagcgt
     1861 cgtcgtagta gttcactgcc gtgtaggcag ctaagaaaaa gccgggccga tgttcgagcc
     1921 tggctgcgtg gttcactgcc gtgtaggcag ctaagaaatg gacaatgcgc agcccaggcg
     1981 ctggatttac gttcactgcc gtgtaggcag ctaagaaaat gcgaggctga tcgaggtgcg
     2041 cggttctgtt gttcactgcc gtgtaggcag ctaagaaagt gcgcggatgg ctggtatctc
     2101 gagcacgatt gttcactgcc gtgtaggcag ctaagaaaag gcaaacaacg gaaagatcac
     2161 caccgaaggc gttcactgcc gtgtaggcag ctaagaaaaa cgaaacaggc ccagcacagc
     2221 cgcttgataa gttcactgcc gtgtaggcag ctaagaaaac aagacgctca tcgacggcag
     2281 catcgagacg gttcactgcc gtgtaggcag ctaagaaaaa ggtgacgatg cacagctgtt
     2341 gcgcgcggtt gttcactgcc gtgtaggcag ctaagaaaat cctcgaacag cccgggttcg
     2401 gtcgcctcca gttcactgcc gtgtaggcag ctaagaaaag gagtttgcgg ccccgctccc
     2461 tgcggctctc gttcactgcc gtgtaggcag ctaagaaatt cacggcgggc ttgatgtccg
     2521 cgtctacctg gttcactgcc gtgtaggcag ctaagaaatt gccgagtacg atgcctgata
     2581 catgaatcca gttcactgcc gtgtaggcag ctaagaaact cgaacccacc tcggccacaa
     2641 cagccgccgg gttcgctgcc gtctaggcag aaccaccctc cccatcccac taccaaacat
     2701 ccgaatataa agttcctacc ccgcccgcca gcctcgcccg tccacgacaa tgtgcctcgc
     2761 ttggaagctc acgctcctca cacagacgaa aacggggtag cggcaatcgg ccatatccgc
     2821 taaacagttg ccttggcgca gaattcgata gatccgatag ggacaggcca cggtcaacat
     2881 ggacgacatt tctcccagcg aactgaagac gatccttcac tcgcatgcaa gcttggcact
     2941 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3001 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3061 ttcccaacag ttgcgcagcc tgaaaggcag gccgggccgt ggtggccacg gcctctaggc
     3121 cagatccagc ggcatctggg ttagtcgagc gcgggccgct tcccatgtct caccagggcg
     3181 agcctgtttc gcgatctcag catctgaaat cttcccggcc ttgcgcttcg ctggggcctt
     3241 acccaccgcc ttggcgggct tcttcggtcc aaaactgaac aacagatgtg tgaccttgcg
     3301 cccggtcttt cgctgcgccc actccacctg tagcgggctg tgctcgttga tctgcgtcac
     3361 ggctggatca agcactcgca acttgaagtc cttgatcgag ggataccggc cttccagttg
     3421 aaaccacttt cgcagctggt caatttctat ttcgcgctgg ccgatgctgt cccattgcat
     3481 gagcagctcg taaagcctga tcgcgtgggt gctgtccatc ttggccacgt cagccaaggc
     3541 gtatttggtg aactgtttgg tgagttccgt caggtacggc agcatgtctt tggtgaacct
     3601 gagttctaca cggccctcac cctcccggta gatgattgtt tgcacccagc cggtaatcat
     3661 cacactcggt cttttcccct tgccattggg ctcttgggtt aaccggactt cccgccgttt
     3721 caggcgcagg gccgcttctt tgagctggtt gtaggaagat tcgataggga cacccgccat
     3781 cgtcgctatg tcctccgccg tcactgaata catcacttca tcggtgacag gctcgctcct
     3841 cttcacctgg ctaatacagg ccagaacgat ccgctgttcc tgaacactga ggcgatacgc
     3901 ggcctcgacc agggcattgc ttttgtaaac cattgggggt gaggccacgt tcgacattcc
     3961 ttgtgtataa ggggacactg tatctgcgtc ccacaataca acaaatccgt ccctttacaa
     4021 caacaaatcc gtcccttctt aacaacaaat ccgtccctta atggcaacaa atccgtccct
     4081 ttttaaactc tacaggccac ggattacgtg gcctgtagac gtcctaaaag gtttaaaagg
     4141 gaaaaggaag aaaagggtgg aaacgcaaaa aacgcaccac tacgtggccc cgttggggcc
     4201 gcatttgtgc ccctgaaggg gcgggggagg cgtctgggca atccccgttt taccagtccc
     4261 ctatcgccgc ctgagagggc gcaggaagcg agtaatcagg gtatcgaggc ggattcaccc
     4321 ttggcgtcca accagcggca ccagcggcgc gaattgacat aagcctgttc ggttcgtaaa
     4381 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     4441 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tgtacagtct
     4501 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     4561 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt
     4621 atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct
     4681 cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac
     4741 tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac
     4801 caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt
     4861 agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc
     4921 accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac
     4981 gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg
     5041 gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc
     5101 gagatcggct tcccggccgc ggagttgttc ggtaaattgt cacaacgccg cgtccaattc
     5161 gactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5221 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5281 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5341 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5401 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5461 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5521 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5581 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5641 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5701 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5761 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5821 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5881 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5941 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6001 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6061 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca caagctagct
     6541 tatcggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac
     6601 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
     6661 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
     6721 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
     6781 ggaaagtata ccttaaggaa tccccaaatt atgacaactt gacggctaca tcattcactt
     6841 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     6901 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     6961 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     7021 cgctaatccc taactgctgg cgga
//
