LOCUS       sequence-301638-        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 10bp addition downstream,and 
            7bp addition upstream of IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7061)
  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 7061)
  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_+10D+7U
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..303
                     /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     226..245
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             330..352
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2304..2320)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2313..2335)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2481..3314)
                     /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      3328..3679
                     /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(3720..3725,3726..3742,3743..3748)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3918..4451
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      5252..5840
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5741..5760
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            5910..6018
                     /note="incP origin of transfer"
     CDS             complement(6175..7053)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
       61 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
      121 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
      181 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
      241 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
      301 catacccgtt tttttgggct agaaataatt ttgtttaact ttaagaagga gatatacata
      361 cccatggatg ccttgtccga aatcgaggcg ggccacccga gcctgtagcg ccgctccctg
      421 gcgcgacgcc cccggcctgg ccgggggctt tgcggcatcg cccatcaaaa gacctttcgc
      481 gcccgaacgg cacggttgat cgccgtcccg gtcctcgcga aacggccgcc aattgcccga
      541 agcttccgac ccttttttcg gacgatttct tacgccctta taaatcagca agttacgaga
      601 cctcgaaaaa agagggtttc tggcgggata cggcaaaaac tcggtatttc tttttccttc
      661 aaatggttaa aggagctatt aggttttcgg agctagttca ctgccgtgta ggcagctaag
      721 aaaatcagcc ggacgttgta gtagtcgagc gcggtgttca ctgccgtgta ggcagctaag
      781 aaagccgggt ggtgacttct ggacggcgtt cgtcaagttc actgccgtgt aggcagctaa
      841 gaaagacctg gttttggccg acacagtggc tgcgacgttc actgccgtgt aggcagctaa
      901 gaaacggtcc tgccagtact gcggcaggat ctggtcgttc actgccgtgt aggcagctaa
      961 gaaaagcctg gcgcccttcg agtccatcca gcccgggttc actgccgtgt aggcagctaa
     1021 gaaacatcca attcgccgat ccggcgcaag cgcccggttc actgccgtgt aggcagctaa
     1081 gaaaacgaag cagatcgccg aatgcggcgt cagcgcgttc actgccgtgt aggcagctaa
     1141 gaaagtgggt gcaaatcgcg attctggtcg cgtcgtgttc actgccgtgt aggcagctaa
     1201 gaaatgaata tcagcctgca aagcgtcgtc gtagtagttc actgccgtgt aggcagctaa
     1261 gaaaaagccg ggccgatgtt cgagcctggc tgcgtggttc actgccgtgt aggcagctaa
     1321 gaaatggaca atgcgcagcc caggcgctgg atttacgttc actgccgtgt aggcagctaa
     1381 gaaaatgcga ggctgatcga ggtgcgcggt tctgttgttc actgccgtgt aggcagctaa
     1441 gaaagtgcgc ggatggctgg tatctcgagc acgattgttc actgccgtgt aggcagctaa
     1501 gaaaaggcaa acaacggaaa gatcaccacc gaaggcgttc actgccgtgt aggcagctaa
     1561 gaaaaacgaa acaggcccag cacagccgct tgataagttc actgccgtgt aggcagctaa
     1621 gaaaacaaga cgctcatcga cggcagcatc gagacggttc actgccgtgt aggcagctaa
     1681 gaaaaaggtg acgatgcaca gctgttgcgc gcggttgttc actgccgtgt aggcagctaa
     1741 gaaaatcctc gaacagcccg ggttcggtcg cctccagttc actgccgtgt aggcagctaa
     1801 gaaaaggagt ttgcggcccc gctccctgcg gctctcgttc actgccgtgt aggcagctaa
     1861 gaaattcacg gcgggcttga tgtccgcgtc tacctggttc actgccgtgt aggcagctaa
     1921 gaaattgccg agtacgatgc ctgatacatg aatccagttc actgccgtgt aggcagctaa
     1981 gaaactcgaa cccacctcgg ccacaacagc cgccgggttc gctgccgtct aggcagaacc
     2041 accctcccca tcccactacc aaacatccga atataaagtt cctaccccgc ccgccagcct
     2101 cgcccgtcca cgacaatgtg cctcgcttgg aagctcacgc tcctcacaca gacgaaaacg
     2161 gggtagcggc aatcggccat atccgctaaa cagttgcctt ggcgcagaat tcgatagatc
     2221 cgatagggac aggccacggt caacatggac gacatttctc ccagcgaact gaagacgatc
     2281 cttcactcgc atgcaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2341 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2401 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa aggcaggccg
     2461 ggccgtggtg gccacggcct ctaggccaga tccagcggca tctgggttag tcgagcgcgg
     2521 gccgcttccc atgtctcacc agggcgagcc tgtttcgcga tctcagcatc tgaaatcttc
     2581 ccggccttgc gcttcgctgg ggccttaccc accgccttgg cgggcttctt cggtccaaaa
     2641 ctgaacaaca gatgtgtgac cttgcgcccg gtctttcgct gcgcccactc cacctgtagc
     2701 gggctgtgct cgttgatctg cgtcacggct ggatcaagca ctcgcaactt gaagtccttg
     2761 atcgagggat accggccttc cagttgaaac cactttcgca gctggtcaat ttctatttcg
     2821 cgctggccga tgctgtccca ttgcatgagc agctcgtaaa gcctgatcgc gtgggtgctg
     2881 tccatcttgg ccacgtcagc caaggcgtat ttggtgaact gtttggtgag ttccgtcagg
     2941 tacggcagca tgtctttggt gaacctgagt tctacacggc cctcaccctc ccggtagatg
     3001 attgtttgca cccagccggt aatcatcaca ctcggtcttt tccccttgcc attgggctct
     3061 tgggttaacc ggacttcccg ccgtttcagg cgcagggccg cttctttgag ctggttgtag
     3121 gaagattcga tagggacacc cgccatcgtc gctatgtcct ccgccgtcac tgaatacatc
     3181 acttcatcgg tgacaggctc gctcctcttc acctggctaa tacaggccag aacgatccgc
     3241 tgttcctgaa cactgaggcg atacgcggcc tcgaccaggg cattgctttt gtaaaccatt
     3301 gggggtgagg ccacgttcga cattccttgt gtataagggg acactgtatc tgcgtcccac
     3361 aatacaacaa atccgtccct ttacaacaac aaatccgtcc cttcttaaca acaaatccgt
     3421 cccttaatgg caacaaatcc gtcccttttt aaactctaca ggccacggat tacgtggcct
     3481 gtagacgtcc taaaaggttt aaaagggaaa aggaagaaaa gggtggaaac gcaaaaaacg
     3541 caccactacg tggccccgtt ggggccgcat ttgtgcccct gaaggggcgg gggaggcgtc
     3601 tgggcaatcc ccgttttacc agtcccctat cgccgcctga gagggcgcag gaagcgagta
     3661 atcagggtat cgaggcggat tcacccttgg cgtccaacca gcggcaccag cggcgcgaat
     3721 tgacataagc ctgttcggtt cgtaaactgt aatgcaagta gcgtatgcgc tcacgcaact
     3781 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     3841 gttatgactg tttttttgta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     3901 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     3961 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
     4021 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag
     4081 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
     4141 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
     4201 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
     4261 gcgagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
     4321 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
     4381 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
     4441 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
     4501 aattgtcaca acgccgcggc caattcgact caccagtcac agaaaagcat cttacggatg
     4561 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4621 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4681 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4741 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4801 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4861 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4921 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4981 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5041 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5101 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5161 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5221 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5281 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5341 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5401 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5461 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5521 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5581 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5641 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5701 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5761 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5821 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5881 gctggccttt tgctcacaag ctagcttatc ggccagcctc gcagagcagg attcccgttg
     5941 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     6001 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     6061 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     6121 atgaccccga agcagggtta tgcagcggaa agtatacctt aaggaatccc caaattatga
     6181 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     6241 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     6301 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     6361 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga
     6421 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     6481 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     6541 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     6601 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     6661 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     6721 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     6781 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     6841 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     6901 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     6961 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     7021 cggcagcagg ggatcatttt gcgcttcagc catacttttc a
//
