LOCUS       sequence-292026-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 1bp addition downstream of 
            IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  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 7065)
  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_+1D
FEATURES             Location/Qualifiers
     source          1..7065
                     /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(4170..4186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4179..4201)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4347..5180)
                     /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      5194..5545
                     /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(5586..5591,5592..5608,5609..5614)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5803..6336
                     /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 ggttagtagg ttttcggagc tagttcactg ccgtgtaggc
     2581 agctaagaaa atcagccgga cgttgtagta gtcgagcgcg gtgttcactg ccgtgtaggc
     2641 agctaagaaa gccgggtggt gacttctgga cggcgttcgt caagttcact gccgtgtagg
     2701 cagctaagaa agacctggtt ttggccgaca cagtggctgc gacgttcact gccgtgtagg
     2761 cagctaagaa acggtcctgc cagtactgcg gcaggatctg gtcgttcact gccgtgtagg
     2821 cagctaagaa aagcctggcg cccttcgagt ccatccagcc cgggttcact gccgtgtagg
     2881 cagctaagaa acatccaatt cgccgatccg gcgcaagcgc ccggttcact gccgtgtagg
     2941 cagctaagaa aacgaagcag atcgccgaat gcggcgtcag cgcgttcact gccgtgtagg
     3001 cagctaagaa agtgggtgca aatcgcgatt ctggtcgcgt cgtgttcact gccgtgtagg
     3061 cagctaagaa atgaatatca gcctgcaaag cgtcgtcgta gtagttcact gccgtgtagg
     3121 cagctaagaa aaagccgggc cgatgttcga gcctggctgc gtggttcact gccgtgtagg
     3181 cagctaagaa atggacaatg cgcagcccag gcgctggatt tacgttcact gccgtgtagg
     3241 cagctaagaa aatgcgaggc tgatcgaggt gcgcggttct gttgttcact gccgtgtagg
     3301 cagctaagaa agtgcgcgga tggctggtat ctcgagcacg attgttcact gccgtgtagg
     3361 cagctaagaa aaggcaaaca acggaaagat caccaccgaa ggcgttcact gccgtgtagg
     3421 cagctaagaa aaacgaaaca ggcccagcac agccgcttga taagttcact gccgtgtagg
     3481 cagctaagaa aacaagacgc tcatcgacgg cagcatcgag acggttcact gccgtgtagg
     3541 cagctaagaa aaaggtgacg atgcacagct gttgcgcgcg gttgttcact gccgtgtagg
     3601 cagctaagaa aatcctcgaa cagcccgggt tcggtcgcct ccagttcact gccgtgtagg
     3661 cagctaagaa aaggagtttg cggccccgct ccctgcggct ctcgttcact gccgtgtagg
     3721 cagctaagaa attcacggcg ggcttgatgt ccgcgtctac ctggttcact gccgtgtagg
     3781 cagctaagaa attgccgagt acgatgcctg atacatgaat ccagttcact gccgtgtagg
     3841 cagctaagaa actcgaaccc acctcggcca caacagccgc cgggttcgct gccgtctagg
     3901 cagaaccacc ctccccatcc cactaccaaa catccgaata taaagttcct accccgcccg
     3961 ccagcctcgc ccgtccacga caatgtgcct cgcttggaag ctcacgctcc tcacacagac
     4021 gaaaacgggg tagcggcaat cggccatatc cgctaaacag ttgccttggc gcagaattcg
     4081 atagatccga tagggacagg ccacggtcaa catggacgac atttctccca gcgaactgaa
     4141 gacgatcctt cactgcatgc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     4201 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4261 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaaaggc
     4321 aggccgggcc gtggtggcca cggcctctag gccagatcca gcggcatctg ggttagtcga
     4381 gcgcgggccg cttcccatgt ctcaccaggg cgagcctgtt tcgcgatctc agcatctgaa
     4441 atcttcccgg ccttgcgctt cgctggggcc ttacccaccg ccttggcggg cttcttcggt
     4501 ccaaaactga acaacagatg tgtgaccttg cgcccggtct ttcgctgcgc ccactccacc
     4561 tgtagcgggc tgtgctcgtt gatctgcgtc acggctggat caagcactcg caacttgaag
     4621 tccttgatcg agggataccg gccttccagt tgaaaccact ttcgcagctg gtcaatttct
     4681 atttcgcgct ggccgatgct gtcccattgc atgagcagct cgtaaagcct gatcgcgtgg
     4741 gtgctgtcca tcttggccac gtcagccaag gcgtatttgg tgaactgttt ggtgagttcc
     4801 gtcaggtacg gcagcatgtc tttggtgaac ctgagttcta cacggccctc accctcccgg
     4861 tagatgattg tttgcaccca gccggtaatc atcacactcg gtcttttccc cttgccattg
     4921 ggctcttggg ttaaccggac ttcccgccgt ttcaggcgca gggccgcttc tttgagctgg
     4981 ttgtaggaag attcgatagg gacacccgcc atcgtcgcta tgtcctccgc cgtcactgaa
     5041 tacatcactt catcggtgac aggctcgctc ctcttcacct ggctaataca ggccagaacg
     5101 atccgctgtt cctgaacact gaggcgatac gcggcctcga ccagggcatt gcttttgtaa
     5161 accattgggg gtgaggccac gttcgacatt ccttgtgtat aaggggacac tgtatctgcg
     5221 tcccacaata caacaaatcc gtccctttac aacaacaaat ccgtcccttc ttaacaacaa
     5281 atccgtccct taatggcaac aaatccgtcc ctttttaaac tctacaggcc acggattacg
     5341 tggcctgtag acgtcctaaa aggtttaaaa gggaaaagga agaaaagggt ggaaacgcaa
     5401 aaaacgcacc actacgtggc cccgttgggg ccgcatttgt gcccctgaag gggcggggga
     5461 ggcgtctggg caatccccgt tttaccagtc ccctatcgcc gcctgagagg gcgcaggaag
     5521 cgagtaatca gggtatcgag gcggattcac ccttggcgtc caaccagcgg caccagcggc
     5581 gcgaattgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac
     5641 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     5701 tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc
     5761 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagcaac
     5821 gatgttacgc agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc
     5881 attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt
     5941 ttcggtcgtg agttcggaga cgtagccacc tactcccaac atcagccgga ctccgattac
     6001 ctcgggaact tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg
     6061 gttgttggcg ctctcgcggc ttacgttctg cccaggtttg agcagccgcg tagtgagatc
     6121 tatatctatg atctcgcagt ctccggcgag caccggaggc agggcattgc caccgcgctc
     6181 atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca
     6241 gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg
     6301 atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatcggc
     6361 ttcccggccg cggagttgtt cggtaaattg tcacaacgcc gcggccaatt cgactcacca
     6421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatg
//
