LOCUS       sequence-300624-        7049 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 7049)
  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 7049)
  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..7049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     679..701
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     694..710
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             2650..2672
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(2699..2983)
                     /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(2757..2776)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3010..3888
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            4045..4153
                     /note="incP origin of transfer"
     rep_origin      complement(4223..4811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4303..4322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5612..6145)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6315..6320,6321..6337,6338..6343))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      6384..6735
                     /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             join(6749..7049,1..533)
                     /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"
ORIGIN
        1 gggtgcaaac aatcatctac cgggagggtg agggccgtgt agaactcagg ttcaccaaag
       61 acatgctgcc gtacctgacg gaactcacca aacagttcac caaatacgcc ttggctgacg
      121 tggccaagat ggacagcacc cacgcgatca ggctttacga gctgctcatg caatgggaca
      181 gcatcggcca gcgcgaaata gaaattgacc agctgcgaaa gtggtttcaa ctggaaggcc
      241 ggtatccctc gatcaaggac ttcaagttgc gagtgcttga tccagccgtg acgcagatca
      301 acgagcacag cccgctacag gtggagtggg cgcagcgaaa gaccgggcgc aaggtcacac
      361 atctgttgtt cagttttgga ccgaagaagc ccgccaaggc ggtgggtaag gccccagcga
      421 agcgcaaggc cgggaagatt tcagatgctg agatcgcgaa acaggctcgc cctggtgaga
      481 catgggaagc ggcccgcgct cgactaaccc agatgccgct ggatctggcc tagaggccgt
      541 ggccaccacg gcccggcctg cctttcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      601 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      661 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagcttg
      721 catgcgagtg aaggatcgtc ttcagttcgc tgggagaaat gtcgtccatg ttgaccgtgg
      781 cctgtcccta tcggatctat cgaattctgc gccaaggcaa ctgtttagcg gatatggccg
      841 attgccgcta ccccgttttc gtctgtgtga ggagcgtgag cttccaagcg aggcacattg
      901 tcgtggacgg gcgaggctgg cgggcggggt aggaacttta tattcggatg tttggtagtg
      961 ggatggggag ggtggttctg cctagacggc agcgaacccg gcggctgttg tggccgaggt
     1021 gggttcgagt ttcttagctg cctacacggc agtgaactgg attcatgtat caggcatcgt
     1081 actcggcaat ttcttagctg cctacacggc agtgaaccag gtagacgcgg acatcaagcc
     1141 cgccgtgaat ttcttagctg cctacacggc agtgaacgag agccgcaggg agcggggccg
     1201 caaactcctt ttcttagctg cctacacggc agtgaactgg aggcgaccga acccgggctg
     1261 ttcgaggatt ttcttagctg cctacacggc agtgaacaac cgcgcgcaac agctgtgcat
     1321 cgtcaccttt ttcttagctg cctacacggc agtgaaccgt ctcgatgctg ccgtcgatga
     1381 gcgtcttgtt ttcttagctg cctacacggc agtgaactta tcaagcggct gtgctgggcc
     1441 tgtttcgttt ttcttagctg cctacacggc agtgaacgcc ttcggtggtg atctttccgt
     1501 tgtttgcctt ttcttagctg cctacacggc agtgaacaat cgtgctcgag ataccagcca
     1561 tccgcgcact ttcttagctg cctacacggc agtgaacaac agaaccgcgc acctcgatca
     1621 gcctcgcatt ttcttagctg cctacacggc agtgaacgta aatccagcgc ctgggctgcg
     1681 cattgtccat ttcttagctg cctacacggc agtgaaccac gcagccaggc tcgaacatcg
     1741 gcccggcttt ttcttagctg cctacacggc agtgaactac tacgacgacg ctttgcaggc
     1801 tgatattcat ttcttagctg cctacacggc agtgaacacg acgcgaccag aatcgcgatt
     1861 tgcacccact ttcttagctg cctacacggc agtgaacgcg ctgacgccgc attcggcgat
     1921 ctgcttcgtt ttcttagctg cctacacggc agtgaaccgg gcgcttgcgc cggatcggcg
     1981 aattggatgt ttcttagctg cctacacggc agtgaacccg ggctggatgg actcgaaggg
     2041 cgccaggctt ttcttagctg cctacacggc agtgaacgac cagatcctgc cgcagtactg
     2101 gcaggaccgt ttcttagctg cctacacggc agtgaacgtc gcagccactg tgtcggccaa
     2161 aaccaggtct ttcttagctg cctacacggc agtgaacttg acgaacgccg tccagaagtc
     2221 accacccggc tttcttagct gcctacacgg cagtgaacac cgcgctcgac tactacaacg
     2281 tccggctgat tttcttagct gcctacacgg cagtgaacta gctccgaaaa cctaatagct
     2341 aaccatttga aggaaaaaga aataccgagt ttttcccgcc agaaaccctc ttttttcgag
     2401 gtctcgtaac ttgctgattt ataagggcgt aagaaatcgt ccgaaaaaag ggtcggaagc
     2461 ttcgggcaat tggcggccgt ttcgcgagga ccgggacggc gatcaaccgt gccgttcggg
     2521 cgcgaaaggt cttttgatgg gcgatgccgc aaagcccccg gccaggccgg gggcgtcgcg
     2581 ccagggagcg gcgctacagg ctcgggtggc ccgcctcgat ttcggacaag gcatccatgg
     2641 gtatgtatat ctccttctta aagttaaaca aaattatttc tagcccaaaa aaacgggtat
     2701 ggagaaacag tagagagttg cgataaaaag cgtcaggtag gatccgctaa tcttatggat
     2761 aaaaatgcta tggcatagca aagtgtgacg ccgtgcaaat aatcaatgtg gacttttctg
     2821 ccgtgattat agacactttt gttacgcgtt tttgtcatgg ctttggtccc gctttgttac
     2881 agaatgcttt taataagcgg ggttaccggt ttggttagcg agaagagcca gtaaaagacg
     2941 cagtgacggc aatgtctgat gcaatatgga caattggttt cttctctgaa tggcgggagt
     3001 atgaaaagta tggctgaagc gcaaaatgat cccctgctgc cgggatactc gtttaatgcc
     3061 catctggtgg cgggtttaac gccgattgag gccaacggtt atctcgattt ttttatcgac
     3121 cgaccgctgg gaatgaaagg ttatattctc aatctcacca ttcgcggtca gggggtggtg
     3181 aaaaatcagg gacgagaatt tgtttgccga ccgggtgata ttttgctgtt cccgccagga
     3241 gagattcatc actacggtcg tcatccggag gctcgcgaat ggtatcacca gtgggtttac
     3301 tttcgtccgc gcgcctactg gcatgaatgg cttaactggc cgtcaatatt tgccaatacg
     3361 gggttctttc gcccggatga agcgcaccag ccgcatttca gcgacctgtt tgggcaaatc
     3421 attaacgccg ggcaagggga agggcgctat tcggagctgc tggcgataaa tctgcttgag
     3481 caattgttac tgcggcgcat ggaagcgatt aacgagtcgc tccatccacc gatggataat
     3541 cgggtacgcg aggcttgtca gtacatcagc gatcacctgg cagacagcaa ttttgatatc
     3601 gccagcgtcg cacagcatgt ttgcttgtcg ccgtcgcgtc tgtcacatct tttccgccag
     3661 cagttaggga ttagcgtctt aagctggcgc gaggaccaac gtatcagcca ggcgaagctg
     3721 cttttgagca ccacccggat gcctatcgcc accgtcggtc gcaatgttgg ttttgacgat
     3781 caactctatt tctcgcgggt atttaaaaaa tgcaccgggg ccagcccgag cgagttccgt
     3841 gccggttgtg aagaaaaagt gaatgatgta gccgtcaagt tgtcataatt tggggattcc
     3901 ttaaggtata ctttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
     3961 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
     4021 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
     4081 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
     4141 gcgaggctgg ccgataagct agcttgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5521 ctgtgactgg tgagtcgaat tggccgcggc gttgtgacaa tttaccgaac aactccgcgg
     5581 ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa
     5641 agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac
     5701 cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga
     5761 gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat
     5821 agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc
     5881 caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt
     5941 tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac
     6001 gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg
     6061 tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg
     6121 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     6181 tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccgtg aaaaccgcca
     6241 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     6301 gctacttgca ttacagttta cgaaccgaac aggcttatgt caattcgcgc cgctggtgcc
     6361 gctggttgga cgccaagggt gaatccgcct cgataccctg attactcgct tcctgcgccc
     6421 tctcaggcgg cgatagggga ctggtaaaac ggggattgcc cagacgcctc ccccgcccct
     6481 tcaggggcac aaatgcggcc ccaacggggc cacgtagtgg tgcgtttttt gcgtttccac
     6541 ccttttcttc cttttccctt ttaaaccttt taggacgtct acaggccacg taatccgtgg
     6601 cctgtagagt ttaaaaaggg acggatttgt tgccattaag ggacggattt gttgttaaga
     6661 agggacggat ttgttgttgt aaagggacgg atttgttgta ttgtgggacg cagatacagt
     6721 gtccccttat acacaaggaa tgtcgaacgt ggcctcaccc ccaatggttt acaaaagcaa
     6781 tgccctggtc gaggccgcgt atcgcctcag tgttcaggaa cagcggatcg ttctggcctg
     6841 tattagccag gtgaagagga gcgagcctgt caccgatgaa gtgatgtatt cagtgacggc
     6901 ggaggacata gcgacgatgg cgggtgtccc tatcgaatct tcctacaacc agctcaaaga
     6961 agcggccctg cgcctgaaac ggcgggaagt ccggttaacc caagagccca atggcaaggg
     7021 gaaaagaccg agtgtgatga ttaccggct
//
