LOCUS       sequence-301639-        7064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 10bp addition downstream,and 
            10bp addition upstream of IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7064)
  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 7064)
  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+10U
FEATURES             Location/Qualifiers
     source          1..7064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     197..216
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             301..323
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2278..2294)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2287..2309)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2455..3288)
                     /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      3302..3653
                     /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(3694..3699,3700..3716,3717..3722)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3892..4425
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      5226..5814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5715..5734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            5884..5992
                     /note="incP origin of transfer"
     CDS             complement(6149..7027)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(7054..7064,1..274)
                     /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)"
ORIGIN
        1 tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc
       61 aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca
      121 aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac
      181 ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga
      241 cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc tagaaataat
      301 tttgtttaac tttaagaagg agatatacat acccatggat gccttgtccg aaatcgaggc
      361 gggccacccg agcctgtagc gccgctccct ggcgcgacgc ccccggcctg gccgggggct
      421 ttgcggcatc gcccatcaaa agacctttcg cgcccgaacg gcacggttga tcgccgtccc
      481 ggtcctcgcg aaacggccgc caattgcccg aagcttccga cccttttttc ggacgatttc
      541 ttacgccctt ataaatcagc aagttacgag acctcgaaaa aagagggttt ctggcgggaa
      601 aaactcaagg agctatggta tttctttttc cttcaaatgg ttataggagc tattaggttt
      661 tcggagctag ttcactgccg tgtaggcagc taagaaaatc agccggacgt tgtagtagtc
      721 gagcgcggtg ttcactgccg tgtaggcagc taagaaagcc gggtggtgac ttctggacgg
      781 cgttcgtcaa gttcactgcc gtgtaggcag ctaagaaaga cctggttttg gccgacacag
      841 tggctgcgac gttcactgcc gtgtaggcag ctaagaaacg gtcctgccag tactgcggca
      901 ggatctggtc gttcactgcc gtgtaggcag ctaagaaaag cctggcgccc ttcgagtcca
      961 tccagcccgg gttcactgcc gtgtaggcag ctaagaaaca tccaattcgc cgatccggcg
     1021 caagcgcccg gttcactgcc gtgtaggcag ctaagaaaac gaagcagatc gccgaatgcg
     1081 gcgtcagcgc gttcactgcc gtgtaggcag ctaagaaagt gggtgcaaat cgcgattctg
     1141 gtcgcgtcgt gttcactgcc gtgtaggcag ctaagaaatg aatatcagcc tgcaaagcgt
     1201 cgtcgtagta gttcactgcc gtgtaggcag ctaagaaaaa gccgggccga tgttcgagcc
     1261 tggctgcgtg gttcactgcc gtgtaggcag ctaagaaatg gacaatgcgc agcccaggcg
     1321 ctggatttac gttcactgcc gtgtaggcag ctaagaaaat gcgaggctga tcgaggtgcg
     1381 cggttctgtt gttcactgcc gtgtaggcag ctaagaaagt gcgcggatgg ctggtatctc
     1441 gagcacgatt gttcactgcc gtgtaggcag ctaagaaaag gcaaacaacg gaaagatcac
     1501 caccgaaggc gttcactgcc gtgtaggcag ctaagaaaaa cgaaacaggc ccagcacagc
     1561 cgcttgataa gttcactgcc gtgtaggcag ctaagaaaac aagacgctca tcgacggcag
     1621 catcgagacg gttcactgcc gtgtaggcag ctaagaaaaa ggtgacgatg cacagctgtt
     1681 gcgcgcggtt gttcactgcc gtgtaggcag ctaagaaaat cctcgaacag cccgggttcg
     1741 gtcgcctcca gttcactgcc gtgtaggcag ctaagaaaag gagtttgcgg ccccgctccc
     1801 tgcggctctc gttcactgcc gtgtaggcag ctaagaaatt cacggcgggc ttgatgtccg
     1861 cgtctacctg gttcactgcc gtgtaggcag ctaagaaatt gccgagtacg atgcctgata
     1921 catgaatcca gttcactgcc gtgtaggcag ctaagaaact cgaacccacc tcggccacaa
     1981 cagccgccgg gttcgctgcc gtctaggcag aaccaccctc cccatcccac taccaaacat
     2041 ccgaatataa agttcctacc ccgcccgcca gcctcgcccg tccacgacaa tgtgcctcgc
     2101 ttggaagctc acgctcctca cacagacgaa aacggggtag cggcaatcgg ccatatccgc
     2161 taaacagttg ccttggcgca gaattcgata gatccgatag ggacaggcca cggtcaacat
     2221 ggacgacatt tctcccagcg aactgaagac gatccttcac tcgcatgcaa gcttggcact
     2281 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2341 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2401 ttcccaacag ttgcgcagcc tgaaaggcag gccgggccgt ggtggccacg gcctctaggc
     2461 cagatccagc ggcatctggg ttagtcgagc gcgggccgct tcccatgtct caccagggcg
     2521 agcctgtttc gcgatctcag catctgaaat cttcccggcc ttgcgcttcg ctggggcctt
     2581 acccaccgcc ttggcgggct tcttcggtcc aaaactgaac aacagatgtg tgaccttgcg
     2641 cccggtcttt cgctgcgccc actccacctg tagcgggctg tgctcgttga tctgcgtcac
     2701 ggctggatca agcactcgca acttgaagtc cttgatcgag ggataccggc cttccagttg
     2761 aaaccacttt cgcagctggt caatttctat ttcgcgctgg ccgatgctgt cccattgcat
     2821 gagcagctcg taaagcctga tcgcgtgggt gctgtccatc ttggccacgt cagccaaggc
     2881 gtatttggtg aactgtttgg tgagttccgt caggtacggc agcatgtctt tggtgaacct
     2941 gagttctaca cggccctcac cctcccggta gatgattgtt tgcacccagc cggtaatcat
     3001 cacactcggt cttttcccct tgccattggg ctcttgggtt aaccggactt cccgccgttt
     3061 caggcgcagg gccgcttctt tgagctggtt gtaggaagat tcgataggga cacccgccat
     3121 cgtcgctatg tcctccgccg tcactgaata catcacttca tcggtgacag gctcgctcct
     3181 cttcacctgg ctaatacagg ccagaacgat ccgctgttcc tgaacactga ggcgatacgc
     3241 ggcctcgacc agggcattgc ttttgtaaac cattgggggt gaggccacgt tcgacattcc
     3301 ttgtgtataa ggggacactg tatctgcgtc ccacaataca acaaatccgt ccctttacaa
     3361 caacaaatcc gtcccttctt aacaacaaat ccgtccctta atggcaacaa atccgtccct
     3421 ttttaaactc tacaggccac ggattacgtg gcctgtagac gtcctaaaag gtttaaaagg
     3481 gaaaaggaag aaaagggtgg aaacgcaaaa aacgcaccac tacgtggccc cgttggggcc
     3541 gcatttgtgc ccctgaaggg gcgggggagg cgtctgggca atccccgttt taccagtccc
     3601 ctatcgccgc ctgagagggc gcaggaagcg agtaatcagg gtatcgaggc ggattcaccc
     3661 ttggcgtcca accagcggca ccagcggcgc gaattgacat aagcctgttc ggttcgtaaa
     3721 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     3781 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tgtacagtct
     3841 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     3901 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt
     3961 atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct
     4021 cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac
     4081 tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac
     4141 caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt
     4201 agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc
     4261 accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac
     4321 gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg
     4381 gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc
     4441 gagatcggct tcccggccgc ggagttgttc ggtaaattgt cacaacgccg cggccaattc
     4501 gactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4561 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4621 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4681 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4741 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4801 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4861 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4921 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4981 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5041 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5101 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5161 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5341 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca caagctagct
     5881 tatcggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac
     5941 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
     6001 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
     6061 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
     6121 ggaaagtata ccttaaggaa tccccaaatt atgacaactt gacggctaca tcattcactt
     6181 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     6241 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     6301 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     6361 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     6421 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     6481 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     6541 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     6601 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     6661 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     6721 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     6781 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     6841 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     6901 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     6961 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     7021 cagccatact tttcatactc ccgccattca gagaagaaac caat
//
