LOCUS       sequence-300508-        7038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 6bp deletion downstream of 
            IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  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 7038)
  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_-6D
FEATURES             Location/Qualifiers
     source          1..7038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(508..513,514..530,531..536))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      577..928
                     /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             942..1775
                     /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     1921..1943
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1936..1952
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             3881..3903
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(3930..4214)
                     /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(3988..4007)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4241..5119
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            5276..5384
                     /note="incP origin of transfer"
     rep_origin      complement(5454..6042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5534..5553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6843..7038,1..338))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 actgcgagat catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa
       61 gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta
      121 ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg
      181 tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg
      241 ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga
      301 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
      361 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
      421 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
      481 cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaattcg
      541 cgccgctggt gccgctggtt ggacgccaag ggtgaatccg cctcgatacc ctgattactc
      601 gcttcctgcg ccctctcagg cggcgatagg ggactggtaa aacggggatt gcccagacgc
      661 ctcccccgcc ccttcagggg cacaaatgcg gccccaacgg ggccacgtag tggtgcgttt
      721 tttgcgtttc cacccttttc ttccttttcc cttttaaacc ttttaggacg tctacaggcc
      781 acgtaatccg tggcctgtag agtttaaaaa gggacggatt tgttgccatt aagggacgga
      841 tttgttgtta agaagggacg gatttgttgt tgtaaaggga cggatttgtt gtattgtggg
      901 acgcagatac agtgtcccct tatacacaag gaatgtcgaa cgtggcctca cccccaatgg
      961 tttacaaaag caatgccctg gtcgaggccg cgtatcgcct cagtgttcag gaacagcgga
     1021 tcgttctggc ctgtattagc caggtgaaga ggagcgagcc tgtcaccgat gaagtgatgt
     1081 attcagtgac ggcggaggac atagcgacga tggcgggtgt ccctatcgaa tcttcctaca
     1141 accagctcaa agaagcggcc ctgcgcctga aacggcggga agtccggtta acccaagagc
     1201 ccaatggcaa ggggaaaaga ccgagtgtga tgattaccgg ctgggtgcaa acaatcatct
     1261 accgggaggg tgagggccgt gtagaactca ggttcaccaa agacatgctg ccgtacctga
     1321 cggaactcac caaacagttc accaaatacg ccttggctga cgtggccaag atggacagca
     1381 cccacgcgat caggctttac gagctgctca tgcaatggga cagcatcggc cagcgcgaaa
     1441 tagaaattga ccagctgcga aagtggtttc aactggaagg ccggtatccc tcgatcaagg
     1501 acttcaagtt gcgagtgctt gatccagccg tgacgcagat caacgagcac agcccgctac
     1561 aggtggagtg ggcgcagcga aagaccgggc gcaaggtcac acatctgttg ttcagttttg
     1621 gaccgaagaa gcccgccaag gcggtgggta aggccccagc gaagcgcaag gccgggaaga
     1681 tttcagatgc tgagatcgcg aaacaggctc gccctggtga gacatgggaa gcggcccgcg
     1741 ctcgactaac ccagatgccg ctggatctgg cctagaggcc gtggccacca cggcccggcc
     1801 tgcctttcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     1861 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     1921 cccagtcacg acgttgtaaa acgacggcca gtgccaagct tgcatgcgag tgaaggatcg
     1981 tcttcagttc gctgggagaa atgtcgtcca tgttgaccgt ggcctgtccc tatcggatct
     2041 atcgaattct gcgccaaggc aactgtttag cggatatggc cgattgccgc taccccgttt
     2101 tcgtctgtgt gaggagcgtg agcttccaag cgaggcacat tgtcgtggac gggcgaggct
     2161 ggcgggcggg gtaggaactt tatattcgga tgtttggtag tgggatgggg agggtggttc
     2221 tgcctagacg gcagcgaacc cggcggctgt tgtggccgag gtgggttcga gtttcttagc
     2281 tgcctacacg gcagtgaact ggattcatgt atcaggcatc gtactcggca atttcttagc
     2341 tgcctacacg gcagtgaacc aggtagacgc ggacatcaag cccgccgtga atttcttagc
     2401 tgcctacacg gcagtgaacg agagccgcag ggagcggggc cgcaaactcc ttttcttagc
     2461 tgcctacacg gcagtgaact ggaggcgacc gaacccgggc tgttcgagga ttttcttagc
     2521 tgcctacacg gcagtgaaca accgcgcgca acagctgtgc atcgtcacct ttttcttagc
     2581 tgcctacacg gcagtgaacc gtctcgatgc tgccgtcgat gagcgtcttg ttttcttagc
     2641 tgcctacacg gcagtgaact tatcaagcgg ctgtgctggg cctgtttcgt ttttcttagc
     2701 tgcctacacg gcagtgaacg ccttcggtgg tgatctttcc gttgtttgcc ttttcttagc
     2761 tgcctacacg gcagtgaaca atcgtgctcg agataccagc catccgcgca ctttcttagc
     2821 tgcctacacg gcagtgaaca acagaaccgc gcacctcgat cagcctcgca ttttcttagc
     2881 tgcctacacg gcagtgaacg taaatccagc gcctgggctg cgcattgtcc atttcttagc
     2941 tgcctacacg gcagtgaacc acgcagccag gctcgaacat cggcccggct ttttcttagc
     3001 tgcctacacg gcagtgaact actacgacga cgctttgcag gctgatattc atttcttagc
     3061 tgcctacacg gcagtgaaca cgacgcgacc agaatcgcga tttgcaccca ctttcttagc
     3121 tgcctacacg gcagtgaacg cgctgacgcc gcattcggcg atctgcttcg ttttcttagc
     3181 tgcctacacg gcagtgaacc gggcgcttgc gccggatcgg cgaattggat gtttcttagc
     3241 tgcctacacg gcagtgaacc cgggctggat ggactcgaag ggcgccaggc ttttcttagc
     3301 tgcctacacg gcagtgaacg accagatcct gccgcagtac tggcaggacc gtttcttagc
     3361 tgcctacacg gcagtgaacg tcgcagccac tgtgtcggcc aaaaccaggt ctttcttagc
     3421 tgcctacacg gcagtgaact tgacgaacgc cgtccagaag tcaccacccg gctttcttag
     3481 ctgcctacac ggcagtgaac accgcgctcg actactacaa cgtccggctg attttcttag
     3541 ctgcctacac ggcagtgaac tagctaccta taaccatttg aaggaaaaag aaataccgag
     3601 tttttcccgc cagaaaccct cttttttcga ggtctcgtaa cttgctgatt tataagggcg
     3661 taagaaatcg tccgaaaaaa gggtcggaag cttcgggcaa ttggcggccg tttcgcgagg
     3721 accgggacgg cgatcaaccg tgccgttcgg gcgcgaaagg tcttttgatg ggcgatgccg
     3781 caaagccccc ggccaggccg ggggcgtcgc gccagggagc ggcgctacag gctcgggtgg
     3841 cccgcctcga tttcggacaa ggcatccatg ggtatgtata tctccttctt aaagttaaac
     3901 aaaattattt ctagcccaaa aaaacgggta tggagaaaca gtagagagtt gcgataaaaa
     3961 gcgtcaggta ggatccgcta atcttatgga taaaaatgct atggcatagc aaagtgtgac
     4021 gccgtgcaaa taatcaatgt ggacttttct gccgtgatta tagacacttt tgttacgcgt
     4081 ttttgtcatg gctttggtcc cgctttgtta cagaatgctt ttaataagcg gggttaccgg
     4141 tttggttagc gagaagagcc agtaaaagac gcagtgacgg caatgtctga tgcaatatgg
     4201 acaattggtt tcttctctga atggcgggag tatgaaaagt atggctgaag cgcaaaatga
     4261 tcccctgctg ccgggatact cgtttaatgc ccatctggtg gcgggtttaa cgccgattga
     4321 ggccaacggt tatctcgatt tttttatcga ccgaccgctg ggaatgaaag gttatattct
     4381 caatctcacc attcgcggtc agggggtggt gaaaaatcag ggacgagaat ttgtttgccg
     4441 accgggtgat attttgctgt tcccgccagg agagattcat cactacggtc gtcatccgga
     4501 ggctcgcgaa tggtatcacc agtgggttta ctttcgtccg cgcgcctact ggcatgaatg
     4561 gcttaactgg ccgtcaatat ttgccaatac ggggttcttt cgcccggatg aagcgcacca
     4621 gccgcatttc agcgacctgt ttgggcaaat cattaacgcc gggcaagggg aagggcgcta
     4681 ttcggagctg ctggcgataa atctgcttga gcaattgtta ctgcggcgca tggaagcgat
     4741 taacgagtcg ctccatccac cgatggataa tcgggtacgc gaggcttgtc agtacatcag
     4801 cgatcacctg gcagacagca attttgatat cgccagcgtc gcacagcatg tttgcttgtc
     4861 gccgtcgcgt ctgtcacatc ttttccgcca gcagttaggg attagcgtct taagctggcg
     4921 cgaggaccaa cgtatcagcc aggcgaagct gcttttgagc accacccgga tgcctatcgc
     4981 caccgtcggt cgcaatgttg gttttgacga tcaactctat ttctcgcggg tatttaaaaa
     5041 atgcaccggg gccagcccga gcgagttccg tgccggttgt gaagaaaaag tgaatgatgt
     5101 agccgtcaag ttgtcataat ttggggattc cttaaggtat actttccgct gcataaccct
     5161 gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag
     5221 gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca
     5281 ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc
     5341 acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccgataagc tagcttgtga
     5401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5641 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5881 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6061 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtcgaa ttggccgcgg
     6781 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     6841 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     6901 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     6961 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     7021 ctccggtgct cgccggag
//
