LOCUS       sequence-300626-        7054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with 10bp addition downstream of 
            IHFprox site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  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 7054)
  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
FEATURES             Location/Qualifiers
     source          1..7054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(54..932)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        959..1243
                     /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     1166..1185
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1270..1292
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3237..3253)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3246..3268)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3414..4247)
                     /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      4261..4612
                     /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(4653..4658,4659..4675,4676..4681)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4851..5384
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            6843..6951
                     /note="incP origin of transfer"
ORIGIN
        1 tgaccccgaa gcagggttat gcagcggaaa gtatacctta aggaatcccc aaattatgac
       61 aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg
      121 gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg
      181 cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata
      241 cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac
      301 agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct
      361 gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg
      421 agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc
      481 agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg
      541 ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac
      601 ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac
      661 cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa
      721 atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga
      781 atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga
      841 taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc
      901 ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa
      961 gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc
     1021 tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa
     1081 gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat
     1141 tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat
     1201 cctacctgac gctttttatc gcaactctct actgtttctc catacccgtt tttttgggct
     1261 agaaataatt ttgtttaact ttaagaagga gatatacata cccatggatg ccttgtccga
     1321 aatcgaggcg ggccacccga gcctgtagcg ccgctccctg gcgcgacgcc cccggcctgg
     1381 ccgggggctt tgcggcatcg cccatcaaaa gacctttcgc gcccgaacgg cacggttgat
     1441 cgccgtcccg gtcctcgcga aacggccgcc aattgcccga agcttccgac ccttttttcg
     1501 gacgatttct tacgccctta taaatcagca agttacgaga cctcgaaaaa agagggtttc
     1561 tggcgggaaa aactcggtat ttctttttcc ttcaaatggt taaaggagct attaggtttt
     1621 cggagctagt tcactgccgt gtaggcagct aagaaaatca gccggacgtt gtagtagtcg
     1681 agcgcggtgt tcactgccgt gtaggcagct aagaaagccg ggtggtgact tctggacggc
     1741 gttcgtcaag ttcactgccg tgtaggcagc taagaaagac ctggttttgg ccgacacagt
     1801 ggctgcgacg ttcactgccg tgtaggcagc taagaaacgg tcctgccagt actgcggcag
     1861 gatctggtcg ttcactgccg tgtaggcagc taagaaaagc ctggcgccct tcgagtccat
     1921 ccagcccggg ttcactgccg tgtaggcagc taagaaacat ccaattcgcc gatccggcgc
     1981 aagcgcccgg ttcactgccg tgtaggcagc taagaaaacg aagcagatcg ccgaatgcgg
     2041 cgtcagcgcg ttcactgccg tgtaggcagc taagaaagtg ggtgcaaatc gcgattctgg
     2101 tcgcgtcgtg ttcactgccg tgtaggcagc taagaaatga atatcagcct gcaaagcgtc
     2161 gtcgtagtag ttcactgccg tgtaggcagc taagaaaaag ccgggccgat gttcgagcct
     2221 ggctgcgtgg ttcactgccg tgtaggcagc taagaaatgg acaatgcgca gcccaggcgc
     2281 tggatttacg ttcactgccg tgtaggcagc taagaaaatg cgaggctgat cgaggtgcgc
     2341 ggttctgttg ttcactgccg tgtaggcagc taagaaagtg cgcggatggc tggtatctcg
     2401 agcacgattg ttcactgccg tgtaggcagc taagaaaagg caaacaacgg aaagatcacc
     2461 accgaaggcg ttcactgccg tgtaggcagc taagaaaaac gaaacaggcc cagcacagcc
     2521 gcttgataag ttcactgccg tgtaggcagc taagaaaaca agacgctcat cgacggcagc
     2581 atcgagacgg ttcactgccg tgtaggcagc taagaaaaag gtgacgatgc acagctgttg
     2641 cgcgcggttg ttcactgccg tgtaggcagc taagaaaatc ctcgaacagc ccgggttcgg
     2701 tcgcctccag ttcactgccg tgtaggcagc taagaaaagg agtttgcggc cccgctccct
     2761 gcggctctcg ttcactgccg tgtaggcagc taagaaattc acggcgggct tgatgtccgc
     2821 gtctacctgg ttcactgccg tgtaggcagc taagaaattg ccgagtacga tgcctgatac
     2881 atgaatccag ttcactgccg tgtaggcagc taagaaactc gaacccacct cggccacaac
     2941 agccgccggg ttcgctgccg tctaggcaga accaccctcc ccatcccact accaaacatc
     3001 cgaatataaa gttcctaccc cgcccgccag cctcgcccgt ccacgacaat gtgcctcgct
     3061 tggaagctca cgctcctcac acagacgaaa acggggtagc ggcaatcggc catatccgct
     3121 aaacagttgc cttggcgcag aattcgatag atccgatagg gacaggccac ggtcaacatg
     3181 gacgacattt ctcccagcga actgaagacg atccttcact cgcatgcaag cttggcactg
     3241 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     3301 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3361 tcccaacagt tgcgcagcct gaaaggcagg ccgggccgtg gtggccacgg cctctaggcc
     3421 agatccagcg gcatctgggt tagtcgagcg cgggccgctt cccatgtctc accagggcga
     3481 gcctgtttcg cgatctcagc atctgaaatc ttcccggcct tgcgcttcgc tggggcctta
     3541 cccaccgcct tggcgggctt cttcggtcca aaactgaaca acagatgtgt gaccttgcgc
     3601 ccggtctttc gctgcgccca ctccacctgt agcgggctgt gctcgttgat ctgcgtcacg
     3661 gctggatcaa gcactcgcaa cttgaagtcc ttgatcgagg gataccggcc ttccagttga
     3721 aaccactttc gcagctggtc aatttctatt tcgcgctggc cgatgctgtc ccattgcatg
     3781 agcagctcgt aaagcctgat cgcgtgggtg ctgtccatct tggccacgtc agccaaggcg
     3841 tatttggtga actgtttggt gagttccgtc aggtacggca gcatgtcttt ggtgaacctg
     3901 agttctacac ggccctcacc ctcccggtag atgattgttt gcacccagcc ggtaatcatc
     3961 acactcggtc ttttcccctt gccattgggc tcttgggtta accggacttc ccgccgtttc
     4021 aggcgcaggg ccgcttcttt gagctggttg taggaagatt cgatagggac acccgccatc
     4081 gtcgctatgt cctccgccgt cactgaatac atcacttcat cggtgacagg ctcgctcctc
     4141 ttcacctggc taatacaggc cagaacgatc cgctgttcct gaacactgag gcgatacgcg
     4201 gcctcgacca gggcattgct tttgtaaacc attgggggtg aggccacgtt cgacattcct
     4261 tgtgtataag gggacactgt atctgcgtcc cacaatacaa caaatccgtc cctttacaac
     4321 aacaaatccg tcccttctta acaacaaatc cgtcccttaa tggcaacaaa tccgtccctt
     4381 tttaaactct acaggccacg gattacgtgg cctgtagacg tcctaaaagg tttaaaaggg
     4441 aaaaggaaga aaagggtgga aacgcaaaaa acgcaccact acgtggcccc gttggggccg
     4501 catttgtgcc cctgaagggg cgggggaggc gtctgggcaa tccccgtttt accagtcccc
     4561 tatcgccgcc tgagagggcg caggaagcga gtaatcaggg tatcgaggcg gattcaccct
     4621 tggcgtccaa ccagcggcac cagcggcgcg aattgacata agcctgttcg gttcgtaaac
     4681 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     4741 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta
     4801 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     4861 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaggt ggctcaagta
     4921 tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt caaatccatg cgggctgctc
     4981 ttgatctttt cggtcgtgag ttcggagacg tagccaccta ctcccaacat cagccggact
     5041 ccgattacct cgggaacttg ctccgtagta agacattcat cgcgcttgct gccttcgacc
     5101 aagaagcggt tgttggcgct ctcgcggctt acgttctgcc caagtttgag cagccgcgta
     5161 gtgagatcta tatctatgat ctcgcagtct ccggcgagca ccggaggcag ggcattgcca
     5221 ccgcgctcat caatctcctc aagcatgagg ccaacgcgct tggtgcttat gtgatctacg
     5281 tgcaagcaga ttacggtgac gatcccgcag tggctctcta tacaaagttg ggcatacggg
     5341 aagaagtgat gcactttgat atcgacccaa gtaccgccac ctaacaattc gttcaagccg
     5401 agatcggctt cccggccgcg gagttgttcg gtaaattgtc acaacgccgc ggccaattcg
     5461 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5521 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5581 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5641 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5701 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5761 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5821 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5881 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5941 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6001 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac aagctagctt
     6841 atcggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca
     6901 gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc
     6961 cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg
     7021 aaaaaggatg gatataccga aaaaatcgct ataa
//
