LOCUS       sequence-301658-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. aeruginosa PA14 CRISPR2 locus, with a scrambled Upstream 
            recognition motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  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 7044)
  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_motif_scram
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        843..1127
                     /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     1050..1069
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1154..1176
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3111..3127)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3120..3142)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3288..4121)
                     /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      4135..4486
                     /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(4527..4532,4533..4549,4550..4555)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4725..5258
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      6059..6647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6548..6567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            6717..6825
                     /note="incP origin of transfer"
     CDS             complement(join(6982..7044,1..816))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
       61 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
      121 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
      181 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
      241 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
      301 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
      361 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
      421 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
      481 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
      541 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
      601 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
      661 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
      721 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
      781 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag
      841 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
      901 cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
      961 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
     1021 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
     1081 ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgtttttttg
     1141 ggctagaaat aattttgttt aactttaaga aggagatata catacccatg gatgccttgt
     1201 ccgaaatcga ggcgggccac ccgagcctgt agcgccgctc cctggcgcga cgcccccggc
     1261 ctggccgggg gctttgcggc atcgcccatc aaaagacctt tcgcgcccga acggcacggt
     1321 tgatcgccgt cccggtcctc gcgaaacggc cgccaattgc ccgaagcttc cgaccctttt
     1381 ttcggacgat ttcttacgcc cttataaatc agcaagttac gagacctcga taccagtgta
     1441 tttctggcgg gaaaaactcg gtatttcttt ttccttcaaa tggttatagg ttttcggagc
     1501 tagttcactg ccgtgtaggc agctaagaaa atcagccgga cgttgtagta gtcgagcgcg
     1561 gtgttcactg ccgtgtaggc agctaagaaa gccgggtggt gacttctgga cggcgttcgt
     1621 caagttcact gccgtgtagg cagctaagaa agacctggtt ttggccgaca cagtggctgc
     1681 gacgttcact gccgtgtagg cagctaagaa acggtcctgc cagtactgcg gcaggatctg
     1741 gtcgttcact gccgtgtagg cagctaagaa aagcctggcg cccttcgagt ccatccagcc
     1801 cgggttcact gccgtgtagg cagctaagaa acatccaatt cgccgatccg gcgcaagcgc
     1861 ccggttcact gccgtgtagg cagctaagaa aacgaagcag atcgccgaat gcggcgtcag
     1921 cgcgttcact gccgtgtagg cagctaagaa agtgggtgca aatcgcgatt ctggtcgcgt
     1981 cgtgttcact gccgtgtagg cagctaagaa atgaatatca gcctgcaaag cgtcgtcgta
     2041 gtagttcact gccgtgtagg cagctaagaa aaagccgggc cgatgttcga gcctggctgc
     2101 gtggttcact gccgtgtagg cagctaagaa atggacaatg cgcagcccag gcgctggatt
     2161 tacgttcact gccgtgtagg cagctaagaa aatgcgaggc tgatcgaggt gcgcggttct
     2221 gttgttcact gccgtgtagg cagctaagaa agtgcgcgga tggctggtat ctcgagcacg
     2281 attgttcact gccgtgtagg cagctaagaa aaggcaaaca acggaaagat caccaccgaa
     2341 ggcgttcact gccgtgtagg cagctaagaa aaacgaaaca ggcccagcac agccgcttga
     2401 taagttcact gccgtgtagg cagctaagaa aacaagacgc tcatcgacgg cagcatcgag
     2461 acggttcact gccgtgtagg cagctaagaa aaaggtgacg atgcacagct gttgcgcgcg
     2521 gttgttcact gccgtgtagg cagctaagaa aatcctcgaa cagcccgggt tcggtcgcct
     2581 ccagttcact gccgtgtagg cagctaagaa aaggagtttg cggccccgct ccctgcggct
     2641 ctcgttcact gccgtgtagg cagctaagaa attcacggcg ggcttgatgt ccgcgtctac
     2701 ctggttcact gccgtgtagg cagctaagaa attgccgagt acgatgcctg atacatgaat
     2761 ccagttcact gccgtgtagg cagctaagaa actcgaaccc acctcggcca caacagccgc
     2821 cgggttcgct gccgtctagg cagaaccacc ctccccatcc cactaccaaa catccgaata
     2881 taaagttcct accccgcccg ccagcctcgc ccgtccacga caatgtgcct cgcttggaag
     2941 ctcacgctcc tcacacagac gaaaacgggg tagcggcaat cggccatatc cgctaaacag
     3001 ttgccttggc gcagaattcg atagatccga tagggacagg ccacggtcaa catggacgac
     3061 atttctccca gcgaactgaa gacgatcctt cactcgcatg caagcttggc actggccgtc
     3121 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     3181 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3241 cagttgcgca gcctgaaagg caggccgggc cgtggtggcc acggcctcta ggccagatcc
     3301 agcggcatct gggttagtcg agcgcgggcc gcttcccatg tctcaccagg gcgagcctgt
     3361 ttcgcgatct cagcatctga aatcttcccg gccttgcgct tcgctggggc cttacccacc
     3421 gccttggcgg gcttcttcgg tccaaaactg aacaacagat gtgtgacctt gcgcccggtc
     3481 tttcgctgcg cccactccac ctgtagcggg ctgtgctcgt tgatctgcgt cacggctgga
     3541 tcaagcactc gcaacttgaa gtccttgatc gagggatacc ggccttccag ttgaaaccac
     3601 tttcgcagct ggtcaatttc tatttcgcgc tggccgatgc tgtcccattg catgagcagc
     3661 tcgtaaagcc tgatcgcgtg ggtgctgtcc atcttggcca cgtcagccaa ggcgtatttg
     3721 gtgaactgtt tggtgagttc cgtcaggtac ggcagcatgt ctttggtgaa cctgagttct
     3781 acacggccct caccctcccg gtagatgatt gtttgcaccc agccggtaat catcacactc
     3841 ggtcttttcc ccttgccatt gggctcttgg gttaaccgga cttcccgccg tttcaggcgc
     3901 agggccgctt ctttgagctg gttgtaggaa gattcgatag ggacacccgc catcgtcgct
     3961 atgtcctccg ccgtcactga atacatcact tcatcggtga caggctcgct cctcttcacc
     4021 tggctaatac aggccagaac gatccgctgt tcctgaacac tgaggcgata cgcggcctcg
     4081 accagggcat tgcttttgta aaccattggg ggtgaggcca cgttcgacat tccttgtgta
     4141 taaggggaca ctgtatctgc gtcccacaat acaacaaatc cgtcccttta caacaacaaa
     4201 tccgtccctt cttaacaaca aatccgtccc ttaatggcaa caaatccgtc cctttttaaa
     4261 ctctacaggc cacggattac gtggcctgta gacgtcctaa aaggtttaaa agggaaaagg
     4321 aagaaaaggg tggaaacgca aaaaacgcac cactacgtgg ccccgttggg gccgcatttg
     4381 tgcccctgaa ggggcggggg aggcgtctgg gcaatccccg ttttaccagt cccctatcgc
     4441 cgcctgagag ggcgcaggaa gcgagtaatc agggtatcga ggcggattca cccttggcgt
     4501 ccaaccagcg gcaccagcgg cgcgaattga cataagcctg ttcggttcgt aaactgtaat
     4561 gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
     4621 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag tctatgcctc
     4681 gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca
     4741 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca
     4801 tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc
     4861 ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt
     4921 acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag
     4981 cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga
     5041 tctatatcta tgatctcgca gtctccggcg agcaccggag gcagggcatt gccaccgcgc
     5101 tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag
     5161 cagattacgg tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag
     5221 tgatgcactt tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg
     5281 gcttcccggc cgcggagttg ttcggtaaat tgtcacaacg ccgcggccaa ttcgactcac
     5341 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5401 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5461 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5521 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5581 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5641 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5701 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5761 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5821 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5881 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5941 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6001 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6061 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6121 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6181 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6241 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6301 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6361 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6421 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6481 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6541 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6601 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6661 cggccttttt acggttcctg gccttttgct ggccttttgc tcacaagcta gcttatcggc
     6721 cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag
     6781 aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg
     6841 atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag
     6901 gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaagt
     6961 ataccttaag gaatccccaa attatgacaa cttgacggct acatcattca ctttttcttc
     7021 acaaccggca cggaactcgc tcgg
//
