LOCUS       sequence-300623-        7242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 1bp addition downstream of 
            IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7242)
  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 7242)
  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_+1D
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1874..1896
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1923..2207)
                     /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     complement(1981..2000)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2234..3112
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            3269..3377
                     /note="incP origin of transfer"
     rep_origin      complement(3447..4035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3527..3546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4836..5369)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5539..5544,5545..5561,5562..5567))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      5608..5959
                     /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)"
     CDS             5973..6806
                     /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"
     primer_bind     6952..6974
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6967..6983
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gggtggttct gcctagacgg cagcgaaccc ggcggctgtt gtggccgagg tgggttcgag
       61 tttcttagct gcctacacgg cagtgaactt gacgaacgcc gtccagaagt caccacccgg
      121 ctttcttagc tgcctacacg gcagtgaaca ccgcgctcga ctactacaac gtccggctga
      181 ttttcttagc tgcctacacg gcagtgaact agctccgaaa acctaccggc ggctgttgtg
      241 gccgaggtgg gttcgagttt cttagctgcc tacacggcag tgaactggat tcatgtatca
      301 ggcatcgtac tcggcaattt cttagctgcc tacacggcag tgaaccaggt agacgcggac
      361 atcaagcccg ccgtgaattt cttagctgcc tacacggcag tgaacgagag ccgcagggag
      421 cggggccgca aactcctttt cttagctgcc tacacggcag tgaactggag gcgaccgaac
      481 ccgggctgtt cgaggatttt cttagctgcc tacacggcag tgaacaaccg cgcgcaacag
      541 ctgtgcatcg tcaccttttt cttagctgcc tacacggcag tgaaccgtct cgatgctgcc
      601 gtcgatgagc gtcttgtttt cttagctgcc tacacggcag tgaacttatc aagcggctgt
      661 gctgggcctg tttcgttttt cttagctgcc tacacggcag tgaacgcctt cggtggtgat
      721 ctttccgttg tttgcctttt cttagctgcc tacacggcag tgaacaatcg tgctcgagat
      781 accagccatc cgcgcacttt cttagctgcc tacacggcag tgaacaacag aaccgcgcac
      841 ctcgatcagc ctcgcatttt cttagctgcc tacacggcag tgaacgtaaa tccagcgcct
      901 gggctgcgca ttgtccattt cttagctgcc tacacggcag tgaaccacgc agccaggctc
      961 gaacatcggc ccggcttttt cttagctgcc tacacggcag tgaactacta cgacgacgct
     1021 ttgcaggctg atattcattt cttagctgcc tacacggcag tgaacacgac gcgaccagaa
     1081 tcgcgatttg cacccacttt cttagctgcc tacacggcag tgaacgcgct gacgccgcat
     1141 tcggcgatct gcttcgtttt cttagctgcc tacacggcag tgaaccgggc gcttgcgccg
     1201 gatcggcgaa ttggatgttt cttagctgcc tacacggcag tgaacccggg ctggatggac
     1261 tcgaagggcg ccaggctttt cttagctgcc tacacggcag tgaacgacca gatcctgccg
     1321 cagtactggc aggaccgttt cttagctgcc tacacggcag tgaacgtcgc agccactgtg
     1381 tcggccaaaa ccaggtcttt cttagctgcc tacacggcag tgaacttgac gaacgccgtc
     1441 cagaagtcac cacccggctt tcttagctgc ctacacggca gtgaacaccg cgctcgacta
     1501 ctacaacgtc cggctgattt tcttagctgc ctacacggca gtgaactagc tccgaaaacc
     1561 tactaaccat ttgaaggaaa aagaaatacc gagtttttcc cgccagaaac cctctttttt
     1621 cgaggtctcg taacttgctg atttataagg gcgtaagaaa tcgtccgaaa aaagggtcgg
     1681 aagcttcggg caattggcgg ccgtttcgcg aggaccggga cggcgatcaa ccgtgccgtt
     1741 cgggcgcgaa aggtcttttg atgggcgatg ccgcaaagcc cccggccagg ccgggggcgt
     1801 cgcgccaggg agcggcgcta caggctcggg tggcccgcct cgatttcgga caaggcatcc
     1861 atgggtatgt atatctcctt cttaaagtta aacaaaatta tttctagccc aaaaaaacgg
     1921 gtatggagaa acagtagaga gttgcgataa aaagcgtcag gtaggatccg ctaatcttat
     1981 ggataaaaat gctatggcat agcaaagtgt gacgccgtgc aaataatcaa tgtggacttt
     2041 tctgccgtga ttatagacac ttttgttacg cgtttttgtc atggctttgg tcccgctttg
     2101 ttacagaatg cttttaataa gcggggttac cggtttggtt agcgagaaga gccagtaaaa
     2161 gacgcagtga cggcaatgtc tgatgcaata tggacaattg gtttcttctc tgaatggcgg
     2221 gagtatgaaa agtatggctg aagcgcaaaa tgatcccctg ctgccgggat actcgtttaa
     2281 tgcccatctg gtggcgggtt taacgccgat tgaggccaac ggttatctcg atttttttat
     2341 cgaccgaccg ctgggaatga aaggttatat tctcaatctc accattcgcg gtcagggggt
     2401 ggtgaaaaat cagggacgag aatttgtttg ccgaccgggt gatattttgc tgttcccgcc
     2461 aggagagatt catcactacg gtcgtcatcc ggaggctcgc gaatggtatc accagtgggt
     2521 ttactttcgt ccgcgcgcct actggcatga atggcttaac tggccgtcaa tatttgccaa
     2581 tacggggttc tttcgcccgg atgaagcgca ccagccgcat ttcagcgacc tgtttgggca
     2641 aatcattaac gccgggcaag gggaagggcg ctattcggag ctgctggcga taaatctgct
     2701 tgagcaattg ttactgcggc gcatggaagc gattaacgag tcgctccatc caccgatgga
     2761 taatcgggta cgcgaggctt gtcagtacat cagcgatcac ctggcagaca gcaattttga
     2821 tatcgccagc gtcgcacagc atgtttgctt gtcgccgtcg cgtctgtcac atcttttccg
     2881 ccagcagtta gggattagcg tcttaagctg gcgcgaggac caacgtatca gccaggcgaa
     2941 gctgcttttg agcaccaccc ggatgcctat cgccaccgtc ggtcgcaatg ttggttttga
     3001 cgatcaactc tatttctcgc gggtatttaa aaaatgcacc ggggccagcc cgagcgagtt
     3061 ccgtgccggt tgtgaagaaa aagtgaatga tgtagccgtc aagttgtcat aatttgggga
     3121 ttccttaagg tatactttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt
     3181 cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact
     3241 ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga
     3301 gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg
     3361 ctctgcgagg ctggccgata agctagcttg tgagcaaaag gccagcaaaa ggccaggaac
     3421 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3481 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3541 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3601 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3661 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3721 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3781 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3841 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3901 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3961 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4021 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4081 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4141 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4201 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4261 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4321 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4381 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4441 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4501 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4561 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4621 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4681 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4741 ttttctgtga ctggtgagtc gaattggccg cggcgttgtg acaatttacc gaacaactcc
     4801 gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata
     4861 tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg
     4921 tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg
     4981 aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga
     5041 tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga
     5101 gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc
     5161 aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac
     5221 tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta
     5281 catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc
     5341 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     5401 tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc
     5461 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc
     5521 atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaattc gcgccgctgg
     5581 tgccgctggt tggacgccaa gggtgaatcc gcctcgatac cctgattact cgcttcctgc
     5641 gccctctcag gcggcgatag gggactggta aaacggggat tgcccagacg cctcccccgc
     5701 cccttcaggg gcacaaatgc ggccccaacg gggccacgta gtggtgcgtt ttttgcgttt
     5761 ccaccctttt cttccttttc ccttttaaac cttttaggac gtctacaggc cacgtaatcc
     5821 gtggcctgta gagtttaaaa agggacggat ttgttgccat taagggacgg atttgttgtt
     5881 aagaagggac ggatttgttg ttgtaaaggg acggatttgt tgtattgtgg gacgcagata
     5941 cagtgtcccc ttatacacaa ggaatgtcga acgtggcctc acccccaatg gtttacaaaa
     6001 gcaatgccct ggtcgaggcc gcgtatcgcc tcagtgttca ggaacagcgg atcgttctgg
     6061 cctgtattag ccaggtgaag aggagcgagc ctgtcaccga tgaagtgatg tattcagtga
     6121 cggcggagga catagcgacg atggcgggtg tccctatcga atcttcctac aaccagctca
     6181 aagaagcggc cctgcgcctg aaacggcggg aagtccggtt aacccaagag cccaatggca
     6241 aggggaaaag accgagtgtg atgattaccg gctgggtgca aacaatcatc taccgggagg
     6301 gtgagggccg tgtagaactc aggttcacca aagacatgct gccgtacctg acggaactca
     6361 ccaaacagtt caccaaatac gccttggctg acgtggccaa gatggacagc acccacgcga
     6421 tcaggcttta cgagctgctc atgcaatggg acagcatcgg ccagcgcgaa atagaaattg
     6481 accagctgcg aaagtggttt caactggaag gccggtatcc ctcgatcaag gacttcaagt
     6541 tgcgagtgct tgatccagcc gtgacgcaga tcaacgagca cagcccgcta caggtggagt
     6601 gggcgcagcg aaagaccggg cgcaaggtca cacatctgtt gttcagtttt ggaccgaaga
     6661 agcccgccaa ggcggtgggt aaggccccag cgaagcgcaa ggccgggaag atttcagatg
     6721 ctgagatcgc gaaacaggct cgccctggtg agacatggga agcggcccgc gctcgactaa
     6781 cccagatgcc gctggatctg gcctagaggc cgtggccacc acggcccggc ctgcctttca
     6841 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     6901 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     6961 gacgttgtaa aacgacggcc agtgccaagc ttgcatgcga gtgaaggatc gtcttcagtt
     7021 cgctgggaga aatgtcgtcc atgttgaccg tggcctgtcc ctatcggatc tatcgaattc
     7081 tgcgccaagg caactgttta gcggatatgg ccgattgccg ctaccccgtt ttcgtctgtg
     7141 tgaggagcgt gagcttccaa gcgaggcaca ttgtcgtgga cgggcgaggc tggcgggcgg
     7201 ggtaggaact ttatattcgg atgtttggta gtgggatggg ga
//
