LOCUS       sequence-292025-        7069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 5bp addition downstream of 
            IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7069)
  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 7069)
  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..7069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     561..580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            732..840
                     /note="incP origin of transfer"
     CDS             complement(997..1875)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1902..2186
                     /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     2109..2128
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2213..2235
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4174..4190)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4183..4205)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4351..5184)
                     /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      5198..5549
                     /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(5590..5595,5596..5612,5613..5618)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5807..6340
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
       61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacata
      721 agctagctta tcggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
      781 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
      841 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
      901 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
      961 tatgcagcgg aaagtatacc ttaaggaatc cccaaattat gacaacttga cggctacatc
     1021 attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt
     1081 aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat
     1141 aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca
     1201 gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa
     1261 gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat
     1321 gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc
     1381 ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg
     1441 cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg
     1501 cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc
     1561 atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg
     1621 atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca
     1681 aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat
     1741 ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat
     1801 cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt
     1861 ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata
     1921 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt
     1981 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg
     2041 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac
     2101 actttgctat gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt
     2161 atcgcaactc tctactgttt ctccataccc gtttttttgg gctagaaata attttgttta
     2221 actttaagaa ggagatatac atacccatgg atgccttgtc cgaaatcgag gcgggccacc
     2281 cgagcctgta gcgccgctcc ctggcgcgac gcccccggcc tggccggggg ctttgcggca
     2341 tcgcccatca aaagaccttt cgcgcccgaa cggcacggtt gatcgccgtc ccggtcctcg
     2401 cgaaacggcc gccaattgcc cgaagcttcc gacccttttt tcggacgatt tcttacgccc
     2461 ttataaatca gcaagttacg agacctcgaa aaaagagggt ttctggcggg aaaaactcgg
     2521 tatttctttt tccttcaaat ggttagctat taggttttcg gagctagttc actgccgtgt
     2581 aggcagctaa gaaaatcagc cggacgttgt agtagtcgag cgcggtgttc actgccgtgt
     2641 aggcagctaa gaaagccggg tggtgacttc tggacggcgt tcgtcaagtt cactgccgtg
     2701 taggcagcta agaaagacct ggttttggcc gacacagtgg ctgcgacgtt cactgccgtg
     2761 taggcagcta agaaacggtc ctgccagtac tgcggcagga tctggtcgtt cactgccgtg
     2821 taggcagcta agaaaagcct ggcgcccttc gagtccatcc agcccgggtt cactgccgtg
     2881 taggcagcta agaaacatcc aattcgccga tccggcgcaa gcgcccggtt cactgccgtg
     2941 taggcagcta agaaaacgaa gcagatcgcc gaatgcggcg tcagcgcgtt cactgccgtg
     3001 taggcagcta agaaagtggg tgcaaatcgc gattctggtc gcgtcgtgtt cactgccgtg
     3061 taggcagcta agaaatgaat atcagcctgc aaagcgtcgt cgtagtagtt cactgccgtg
     3121 taggcagcta agaaaaagcc gggccgatgt tcgagcctgg ctgcgtggtt cactgccgtg
     3181 taggcagcta agaaatggac aatgcgcagc ccaggcgctg gatttacgtt cactgccgtg
     3241 taggcagcta agaaaatgcg aggctgatcg aggtgcgcgg ttctgttgtt cactgccgtg
     3301 taggcagcta agaaagtgcg cggatggctg gtatctcgag cacgattgtt cactgccgtg
     3361 taggcagcta agaaaaggca aacaacggaa agatcaccac cgaaggcgtt cactgccgtg
     3421 taggcagcta agaaaaacga aacaggccca gcacagccgc ttgataagtt cactgccgtg
     3481 taggcagcta agaaaacaag acgctcatcg acggcagcat cgagacggtt cactgccgtg
     3541 taggcagcta agaaaaaggt gacgatgcac agctgttgcg cgcggttgtt cactgccgtg
     3601 taggcagcta agaaaatcct cgaacagccc gggttcggtc gcctccagtt cactgccgtg
     3661 taggcagcta agaaaaggag tttgcggccc cgctccctgc ggctctcgtt cactgccgtg
     3721 taggcagcta agaaattcac ggcgggcttg atgtccgcgt ctacctggtt cactgccgtg
     3781 taggcagcta agaaattgcc gagtacgatg cctgatacat gaatccagtt cactgccgtg
     3841 taggcagcta agaaactcga acccacctcg gccacaacag ccgccgggtt cgctgccgtc
     3901 taggcagaac caccctcccc atcccactac caaacatccg aatataaagt tcctaccccg
     3961 cccgccagcc tcgcccgtcc acgacaatgt gcctcgcttg gaagctcacg ctcctcacac
     4021 agacgaaaac ggggtagcgg caatcggcca tatccgctaa acagttgcct tggcgcagaa
     4081 ttcgatagat ccgataggga caggccacgg tcaacatgga cgacatttct cccagcgaac
     4141 tgaagacgat ccttcactgc atgcaagctt ggcactggcc gtcgttttac aacgtcgtga
     4201 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4261 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4321 aggcaggccg ggccgtggtg gccacggcct ctaggccaga tccagcggca tctgggttag
     4381 tcgagcgcgg gccgcttccc atgtctcacc agggcgagcc tgtttcgcga tctcagcatc
     4441 tgaaatcttc ccggccttgc gcttcgctgg ggccttaccc accgccttgg cgggcttctt
     4501 cggtccaaaa ctgaacaaca gatgtgtgac cttgcgcccg gtctttcgct gcgcccactc
     4561 cacctgtagc gggctgtgct cgttgatctg cgtcacggct ggatcaagca ctcgcaactt
     4621 gaagtccttg atcgagggat accggccttc cagttgaaac cactttcgca gctggtcaat
     4681 ttctatttcg cgctggccga tgctgtccca ttgcatgagc agctcgtaaa gcctgatcgc
     4741 gtgggtgctg tccatcttgg ccacgtcagc caaggcgtat ttggtgaact gtttggtgag
     4801 ttccgtcagg tacggcagca tgtctttggt gaacctgagt tctacacggc cctcaccctc
     4861 ccggtagatg attgtttgca cccagccggt aatcatcaca ctcggtcttt tccccttgcc
     4921 attgggctct tgggttaacc ggacttcccg ccgtttcagg cgcagggccg cttctttgag
     4981 ctggttgtag gaagattcga tagggacacc cgccatcgtc gctatgtcct ccgccgtcac
     5041 tgaatacatc acttcatcgg tgacaggctc gctcctcttc acctggctaa tacaggccag
     5101 aacgatccgc tgttcctgaa cactgaggcg atacgcggcc tcgaccaggg cattgctttt
     5161 gtaaaccatt gggggtgagg ccacgttcga cattccttgt gtataagggg acactgtatc
     5221 tgcgtcccac aatacaacaa atccgtccct ttacaacaac aaatccgtcc cttcttaaca
     5281 acaaatccgt cccttaatgg caacaaatcc gtcccttttt aaactctaca ggccacggat
     5341 tacgtggcct gtagacgtcc taaaaggttt aaaagggaaa aggaagaaaa gggtggaaac
     5401 gcaaaaaacg caccactacg tggccccgtt ggggccgcat ttgtgcccct gaaggggcgg
     5461 gggaggcgtc tgggcaatcc ccgttttacc agtcccctat cgccgcctga gagggcgcag
     5521 gaagcgagta atcagggtat cgaggcggat tcacccttgg cgtccaacca gcggcaccag
     5581 cggcgcgaat tgacataagc ctgttcggtt cgtaaactgt aatgcaagta gcgtatgcgc
     5641 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     5701 ttcatggctt gttatgactg tttttttgta cagtctatgc ctcgggcatc caagcagcaa
     5761 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     5821 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaggtggct caagtatggg
     5881 catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg ctgctcttga
     5941 tcttttcggt cgtgagttcg gagacgtagc cacctactcc caacatcagc cggactccga
     6001 ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct tcgaccaaga
     6061 agcggttgtt ggcgctctcg cggcttacgt tctgcccagg tttgagcagc cgcgtagtga
     6121 gatctatatc tatgatctcg cagtctccgg cgagcaccgg aggcagggca ttgccaccgc
     6181 gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga tctacgtgca
     6241 agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca tacgggaaga
     6301 agtgatgcac tttgatatcg acccaagtac cgccacctaa caattcgttc aagccgagat
     6361 cggcttcccg gccgcggagt tgttcggtaa attgtcacaa cgccgcggcc aattcgactc
     6421 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6481 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6541 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6601 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6661 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6721 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6781 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6841 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6901 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6961 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7021 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatg
//
