LOCUS       sequence-416376-        7020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with the IHFdistal binding site 
            removed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7020)
  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 7020)
  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_IHFdist_rm
FEATURES             Location/Qualifiers
     source          1..7020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             22..44
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(71..355)
                     /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(129..148)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             382..1260
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            1417..1525
                     /note="incP origin of transfer"
     rep_origin      complement(1595..2183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1675..1694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2984..3517)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3687..3692,3693..3709,3710..3715))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3756..4107
                     /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             4121..4954
                     /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     5100..5122
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5115..5131
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aggcatccat gggtatgtat atctccttct taaagttaaa caaaattatt tctagcccaa
       61 aaaaacgggt atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct
      121 aatcttatgg ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg
      181 tggacttttc tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc
      241 ccgctttgtt acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc
      301 cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttcttctctg
      361 aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct gccgggatac
      421 tcgtttaatg cccatctggt ggcgggttta acgccgattg aggccaacgg ttatctcgat
      481 ttttttatcg accgaccgct gggaatgaaa ggttatattc tcaatctcac cattcgcggt
      541 cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga tattttgctg
      601 ttcccgccag gagagattca tcactacggt cgtcatccgg aggctcgcga atggtatcac
      661 cagtgggttt actttcgtcc gcgcgcctac tggcatgaat ggcttaactg gccgtcaata
      721 tttgccaata cggggttctt tcgcccggat gaagcgcacc agccgcattt cagcgacctg
      781 tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct attcggagct gctggcgata
      841 aatctgcttg agcaattgtt actgcggcgc atggaagcga ttaacgagtc gctccatcca
      901 ccgatggata atcgggtacg cgaggcttgt cagtacatca gcgatcacct ggcagacagc
      961 aattttgata tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg tctgtcacat
     1021 cttttccgcc agcagttagg gattagcgtc ttaagctggc gcgaggacca acgtatcagc
     1081 caggcgaagc tgcttttgag caccacccgg atgcctatcg ccaccgtcgg tcgcaatgtt
     1141 ggttttgacg atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg ggccagcccg
     1201 agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa gttgtcataa
     1261 tttggggatt ccttaaggta tactttccgc tgcataaccc tgcttcgggg tcattatagc
     1321 gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg
     1381 tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc
     1441 acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg
     1501 gaatcctgct ctgcgaggct ggccgataag ctagcttgtg agcaaaaggc cagcaaaagg
     1561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2101 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2161 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2281 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2641 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2881 taagatgctt ttctgtgact ggtgagtcga attggccgcg gcgttgtgac aatttaccga
     2941 acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg
     3001 ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg
     3061 cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct
     3121 catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga
     3181 ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc agaacgtaag
     3241 ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac
     3301 tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag gtggctacgt
     3361 ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact tggtcagggc
     3421 cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt ttagggcgac
     3481 tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
     3541 gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca
     3601 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
     3661 gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat gtcaattcgc
     3721 gccgctggtg ccgctggttg gacgccaagg gtgaatccgc ctcgataccc tgattactcg
     3781 cttcctgcgc cctctcaggc ggcgataggg gactggtaaa acggggattg cccagacgcc
     3841 tcccccgccc cttcaggggc acaaatgcgg ccccaacggg gccacgtagt ggtgcgtttt
     3901 ttgcgtttcc acccttttct tccttttccc ttttaaacct tttaggacgt ctacaggcca
     3961 cgtaatccgt ggcctgtaga gtttaaaaag ggacggattt gttgccatta agggacggat
     4021 ttgttgttaa gaagggacgg atttgttgtt gtaaagggac ggatttgttg tattgtggga
     4081 cgcagataca gtgtcccctt atacacaagg aatgtcgaac gtggcctcac ccccaatggt
     4141 ttacaaaagc aatgccctgg tcgaggccgc gtatcgcctc agtgttcagg aacagcggat
     4201 cgttctggcc tgtattagcc aggtgaagag gagcgagcct gtcaccgatg aagtgatgta
     4261 ttcagtgacg gcggaggaca tagcgacgat ggcgggtgtc cctatcgaat cttcctacaa
     4321 ccagctcaaa gaagcggccc tgcgcctgaa acggcgggaa gtccggttaa cccaagagcc
     4381 caatggcaag gggaaaagac cgagtgtgat gattaccggc tgggtgcaaa caatcatcta
     4441 ccgggagggt gagggccgtg tagaactcag gttcaccaaa gacatgctgc cgtacctgac
     4501 ggaactcacc aaacagttca ccaaatacgc cttggctgac gtggccaaga tggacagcac
     4561 ccacgcgatc aggctttacg agctgctcat gcaatgggac agcatcggcc agcgcgaaat
     4621 agaaattgac cagctgcgaa agtggtttca actggaaggc cggtatccct cgatcaagga
     4681 cttcaagttg cgagtgcttg atccagccgt gacgcagatc aacgagcaca gcccgctaca
     4741 ggtggagtgg gcgcagcgaa agaccgggcg caaggtcaca catctgttgt tcagttttgg
     4801 accgaagaag cccgccaagg cggtgggtaa ggccccagcg aagcgcaagg ccgggaagat
     4861 ttcagatgct gagatcgcga aacaggctcg ccctggtgag acatgggaag cggcccgcgc
     4921 tcgactaacc cagatgccgc tggatctggc ctagaggccg tggccaccac ggcccggcct
     4981 gcctttcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5041 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5101 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt gcatgcgagt gaaggatcgt
     5161 cttcagttcg ctgggagaaa tgtcgtccat gttgaccgtg gcctgtccct atcggatcta
     5221 tcgaattctg cgccaaggca actgtttagc ggatatggcc gattgccgct accccgtttt
     5281 cgtctgtgtg aggagcgtga gcttccaagc gaggcacatt gtcgtggacg ggcgaggctg
     5341 gcgggcgggg taggaacttt atattcggat gtttggtagt gggatgggga gggtggttct
     5401 gcctagacgg cagcgaaccc ggcggctgtt gtggccgagg tgggttcgag tttcttagct
     5461 gcctacacgg cagtgaactg gattcatgta tcaggcatcg tactcggcaa tttcttagct
     5521 gcctacacgg cagtgaacca ggtagacgcg gacatcaagc ccgccgtgaa tttcttagct
     5581 gcctacacgg cagtgaacga gagccgcagg gagcggggcc gcaaactcct tttcttagct
     5641 gcctacacgg cagtgaactg gaggcgaccg aacccgggct gttcgaggat tttcttagct
     5701 gcctacacgg cagtgaacaa ccgcgcgcaa cagctgtgca tcgtcacctt tttcttagct
     5761 gcctacacgg cagtgaaccg tctcgatgct gccgtcgatg agcgtcttgt tttcttagct
     5821 gcctacacgg cagtgaactt atcaagcggc tgtgctgggc ctgtttcgtt tttcttagct
     5881 gcctacacgg cagtgaacgc cttcggtggt gatctttccg ttgtttgcct tttcttagct
     5941 gcctacacgg cagtgaacaa tcgtgctcga gataccagcc atccgcgcac tttcttagct
     6001 gcctacacgg cagtgaacaa cagaaccgcg cacctcgatc agcctcgcat tttcttagct
     6061 gcctacacgg cagtgaacgt aaatccagcg cctgggctgc gcattgtcca tttcttagct
     6121 gcctacacgg cagtgaacca cgcagccagg ctcgaacatc ggcccggctt tttcttagct
     6181 gcctacacgg cagtgaacta ctacgacgac gctttgcagg ctgatattca tttcttagct
     6241 gcctacacgg cagtgaacac gacgcgacca gaatcgcgat ttgcacccac tttcttagct
     6301 gcctacacgg cagtgaacgc gctgacgccg cattcggcga tctgcttcgt tttcttagct
     6361 gcctacacgg cagtgaaccg ggcgcttgcg ccggatcggc gaattggatg tttcttagct
     6421 gcctacacgg cagtgaaccc gggctggatg gactcgaagg gcgccaggct tttcttagct
     6481 gcctacacgg cagtgaacga ccagatcctg ccgcagtact ggcaggaccg tttcttagct
     6541 gcctacacgg cagtgaacgt cgcagccact gtgtcggcca aaaccaggtc tttcttagct
     6601 gcctacacgg cagtgaactt gacgaacgcc gtccagaagt caccacccgg ctttcttagc
     6661 tgcctacacg gcagtgaaca ccgcgctcga ctactacaac gtccggctga ttttcttagc
     6721 tgcctacacg gcagtgaact agctccgaaa acctataacc atttgaagga aaaagaaata
     6781 ccgagttttt cccgccagaa accctctttt ttcgaggtct cgtagaaatc gtccgaaaaa
     6841 agggtcggaa gcttcgggca attggcggcc gtttcgcgag gaccgggacg gcgatcaacc
     6901 gtgccgttcg ggcgcgaaag gtcttttgat gggcgatgcc gcaaagcccc cggccaggcc
     6961 gggggcgtcg cgccagggag cggcgctaca ggctcgggtg gcccgcctcg atttcggaca
//
