LOCUS       sequence-300509-        7039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 5bp deletion downstream of 
            IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7039)
  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 7039)
  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_-5D
FEATURES             Location/Qualifiers
     source          1..7039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1510..1532
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1559..1843)
                     /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(1617..1636)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1870..2748
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            2905..3013
                     /note="incP origin of transfer"
     rep_origin      complement(3083..3671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3163..3182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4472..5005)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5175..5180,5181..5197,5198..5203))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      5244..5595
                     /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             5609..6442
                     /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     6588..6610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6603..6619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acatcaagcc cgccgtgaat ttcttagctg cctacacggc agtgaacgag agccgcaggg
       61 agcggggccg caaactcctt ttcttagctg cctacacggc agtgaactgg aggcgaccga
      121 acccgggctg ttcgaggatt ttcttagctg cctacacggc agtgaacaac cgcgcgcaac
      181 agctgtgcat cgtcaccttt ttcttagctg cctacacggc agtgaaccgt ctcgatgctg
      241 ccgtcgatga gcgtcttgtt ttcttagctg cctacacggc agtgaactta tcaagcggct
      301 gtgctgggcc tgtttcgttt ttcttagctg cctacacggc agtgaacgcc ttcggtggtg
      361 atctttccgt tgtttgcctt ttcttagctg cctacacggc agtgaacaat cgtgctcgag
      421 ataccagcca tccgcgcact ttcttagctg cctacacggc agtgaacaac agaaccgcgc
      481 acctcgatca gcctcgcatt ttcttagctg cctacacggc agtgaacgta aatccagcgc
      541 ctgggctgcg cattgtccat ttcttagctg cctacacggc agtgaaccac gcagccaggc
      601 tcgaacatcg gcccggcttt ttcttagctg cctacacggc agtgaactac tacgacgacg
      661 ctttgcaggc tgatattcat ttcttagctg cctacacggc agtgaacacg acgcgaccag
      721 aatcgcgatt tgcacccact ttcttagctg cctacacggc agtgaacgcg ctgacgccgc
      781 attcggcgat ctgcttcgtt ttcttagctg cctacacggc agtgaaccgg gcgcttgcgc
      841 cggatcggcg aattggatgt ttcttagctg cctacacggc agtgaacccg ggctggatgg
      901 actcgaaggg cgccaggctt ttcttagctg cctacacggc agtgaacgac cagatcctgc
      961 cgcagtactg gcaggaccgt ttcttagctg cctacacggc agtgaacgtc gcagccactg
     1021 tgtcggccaa aaccaggtct ttcttagctg cctacacggc agtgaacttg acgaacgccg
     1081 tccagaagtc accacccggc tttcttagct gcctacacgg cagtgaacac cgcgctcgac
     1141 tactacaacg tccggctgat tttcttagct gcctacacgg cagtgaacta gctcacctat
     1201 aaccatttga aggaaaaaga aataccgagt ttttcccgcc agaaaccctc ttttttcgag
     1261 gtctcgtaac ttgctgattt ataagggcgt aagaaatcgt ccgaaaaaag ggtcggaagc
     1321 ttcgggcaat tggcggccgt ttcgcgagga ccgggacggc gatcaaccgt gccgttcggg
     1381 cgcgaaaggt cttttgatgg gcgatgccgc aaagcccccg gccaggccgg gggcgtcgcg
     1441 ccagggagcg gcgctacagg ctcgggtggc ccgcctcgat ttcggacaag gcatccatgg
     1501 gtatgtatat ctccttctta aagttaaaca aaattatttc tagcccaaaa aaacgggtat
     1561 ggagaaacag tagagagttg cgataaaaag cgtcaggtag gatccgctaa tcttatggat
     1621 aaaaatgcta tggcatagca aagtgtgacg ccgtgcaaat aatcaatgtg gacttttctg
     1681 ccgtgattat agacactttt gttacgcgtt tttgtcatgg ctttggtccc gctttgttac
     1741 agaatgcttt taataagcgg ggttaccggt ttggttagcg agaagagcca gtaaaagacg
     1801 cagtgacggc aatgtctgat gcaatatgga caattggttt cttctctgaa tggcgggagt
     1861 atgaaaagta tggctgaagc gcaaaatgat cccctgctgc cgggatactc gtttaatgcc
     1921 catctggtgg cgggtttaac gccgattgag gccaacggtt atctcgattt ttttatcgac
     1981 cgaccgctgg gaatgaaagg ttatattctc aatctcacca ttcgcggtca gggggtggtg
     2041 aaaaatcagg gacgagaatt tgtttgccga ccgggtgata ttttgctgtt cccgccagga
     2101 gagattcatc actacggtcg tcatccggag gctcgcgaat ggtatcacca gtgggtttac
     2161 tttcgtccgc gcgcctactg gcatgaatgg cttaactggc cgtcaatatt tgccaatacg
     2221 gggttctttc gcccggatga agcgcaccag ccgcatttca gcgacctgtt tgggcaaatc
     2281 attaacgccg ggcaagggga agggcgctat tcggagctgc tggcgataaa tctgcttgag
     2341 caattgttac tgcggcgcat ggaagcgatt aacgagtcgc tccatccacc gatggataat
     2401 cgggtacgcg aggcttgtca gtacatcagc gatcacctgg cagacagcaa ttttgatatc
     2461 gccagcgtcg cacagcatgt ttgcttgtcg ccgtcgcgtc tgtcacatct tttccgccag
     2521 cagttaggga ttagcgtctt aagctggcgc gaggaccaac gtatcagcca ggcgaagctg
     2581 cttttgagca ccacccggat gcctatcgcc accgtcggtc gcaatgttgg ttttgacgat
     2641 caactctatt tctcgcgggt atttaaaaaa tgcaccgggg ccagcccgag cgagttccgt
     2701 gccggttgtg aagaaaaagt gaatgatgta gccgtcaagt tgtcataatt tggggattcc
     2761 ttaaggtata ctttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
     2821 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
     2881 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
     2941 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
     3001 gcgaggctgg ccgataagct agcttgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3961 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4381 ctgtgactgg tgagtcgaat tggccgcggc gttgtgacaa tttaccgaac aactccgcgg
     4441 ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa
     4501 agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac
     4561 cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga
     4621 gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat
     4681 agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc
     4741 caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt
     4801 tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac
     4861 gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg
     4921 tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg
     4981 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     5041 tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca
     5101 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     5161 gctacttgca ttacagttta cgaaccgaac aggcttatgt caattcgcgc cgctggtgcc
     5221 gctggttgga cgccaagggt gaatccgcct cgataccctg attactcgct tcctgcgccc
     5281 tctcaggcgg cgatagggga ctggtaaaac ggggattgcc cagacgcctc ccccgcccct
     5341 tcaggggcac aaatgcggcc ccaacggggc cacgtagtgg tgcgtttttt gcgtttccac
     5401 ccttttcttc cttttccctt ttaaaccttt taggacgtct acaggccacg taatccgtgg
     5461 cctgtagagt ttaaaaaggg acggatttgt tgccattaag ggacggattt gttgttaaga
     5521 agggacggat ttgttgttgt aaagggacgg atttgttgta ttgtgggacg cagatacagt
     5581 gtccccttat acacaaggaa tgtcgaacgt ggcctcaccc ccaatggttt acaaaagcaa
     5641 tgccctggtc gaggccgcgt atcgcctcag tgttcaggaa cagcggatcg ttctggcctg
     5701 tattagccag gtgaagagga gcgagcctgt caccgatgaa gtgatgtatt cagtgacggc
     5761 ggaggacata gcgacgatgg cgggtgtccc tatcgaatct tcctacaacc agctcaaaga
     5821 agcggccctg cgcctgaaac ggcgggaagt ccggttaacc caagagccca atggcaaggg
     5881 gaaaagaccg agtgtgatga ttaccggctg ggtgcaaaca atcatctacc gggagggtga
     5941 gggccgtgta gaactcaggt tcaccaaaga catgctgccg tacctgacgg aactcaccaa
     6001 acagttcacc aaatacgcct tggctgacgt ggccaagatg gacagcaccc acgcgatcag
     6061 gctttacgag ctgctcatgc aatgggacag catcggccag cgcgaaatag aaattgacca
     6121 gctgcgaaag tggtttcaac tggaaggccg gtatccctcg atcaaggact tcaagttgcg
     6181 agtgcttgat ccagccgtga cgcagatcaa cgagcacagc ccgctacagg tggagtgggc
     6241 gcagcgaaag accgggcgca aggtcacaca tctgttgttc agttttggac cgaagaagcc
     6301 cgccaaggcg gtgggtaagg ccccagcgaa gcgcaaggcc gggaagattt cagatgctga
     6361 gatcgcgaaa caggctcgcc ctggtgagac atgggaagcg gcccgcgctc gactaaccca
     6421 gatgccgctg gatctggcct agaggccgtg gccaccacgg cccggcctgc ctttcaggct
     6481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6601 ttgtaaaacg acggccagtg ccaagcttgc atgcgagtga aggatcgtct tcagttcgct
     6661 gggagaaatg tcgtccatgt tgaccgtggc ctgtccctat cggatctatc gaattctgcg
     6721 ccaaggcaac tgtttagcgg atatggccga ttgccgctac cccgttttcg tctgtgtgag
     6781 gagcgtgagc ttccaagcga ggcacattgt cgtggacggg cgaggctggc gggcggggta
     6841 ggaactttat attcggatgt ttggtagtgg gatggggagg gtggttctgc ctagacggca
     6901 gcgaacccgg cggctgttgt ggccgaggtg ggttcgagtt tcttagctgc ctacacggca
     6961 gtgaactgga ttcatgtatc aggcatcgta ctcggcaatt tcttagctgc ctacacggca
     7021 gtgaaccagg tagacgcgg
//
