LOCUS       sequence-344136-        7820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids bearing protospacer sequences with functional PAM 
            efficiently triggered CRISPR-mediated defense in cells with 
            corresponding spacer sequence; article demonstrates use in Haloferax
            mediterranei.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7820)
  AUTHORS   Lin L, Chen J, Mitra R, Gao Q, Cheng F, Xu T, Zuo Z, Xiang H, Han J
  TITLE     Optimising PHBV biopolymer production in haloarchaea via 
            CRISPRi-mediated redirection of carbon flux.
  JOURNAL   Commun Biol. 2021 Aug 25;4(1):1007. doi: 10.1038/s42003-021-02541-z.
  PUBMED    34433872
REFERENCE   2  (bases 1 to 7820)
  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"; journalName: "Commun 
            Biol."; date: "2021-08-25"; volume: "4(1):1007. doi"; pages: " 
            10.1038/s42003-021-02541-z"
COMMENT     Sequence Label: pWL502-PAM
FEATURES             Location/Qualifiers
     source          1..7820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5148..5165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5319..5907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5399..5418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6078..6869,6870..6938))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6701..6720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6939..7043)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7111..7129
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acctccgggt tctcaaggcg cgggaaggcc gacggtcata cgatgagaca attgcggtgg
       61 tgcttgacgc atatctctct gagaaggtgg attaataaac tatagtcgtg actagcgaac
      121 tagttgaatc aaacgacgcc gggagcacac gagtctgcca ggacttcgct cctatcggcg
      181 tcgaaggatg cttacgcgga cacggcgggc gggtggcccg ttgtcctaac tcaaccaatg
      241 tggttggggc ttatgggatt atcggatttt cacccggcgt tagtgttcgc acaaaagcga
      301 cactaaatgt acgatacgga cgatacgacg gcacacgaac cgactgagaa gcgaagcctt
      361 cgctttgcga ctgacctcga ggcggacgtc tcgcttacag actggaccgc ccttcgggtt
      421 catcgagtaa tcttcgacga gacagagagt caatttgcgc ttctcgaggc actacaaaca
      481 gcggcggacg gtgtcgacat actcgagttt gaacaccgac tgtcgagttg tgggtttgac
      541 gtcgatgggt cagtcgtgaa acttgcacga cgagctgtcg ccggcgagag acgggctctg
      601 tgggacttga ttcggtatgc gcgacggttg ggacatgatg agatgggaca gccacagagc
      661 gccttagagg acgctctcgc gttcactcgt ctattgggct atagcaaccc tcaacagacg
      721 atagcaatcg gtctcaatgc ggctttcaga gagacgcggc gcgaccaggc cgacgagatt
      781 atcgagtacg ttctaacgct gtctgagagc gtcgacgtgg tccttggtgg gtcagagatc
      841 gaccgagtgt acttgcacaa ctactacggc gatattctcc catcgagtgt tacgaatccg
      901 tgcaacgaca cccgtaatga ttactccatt acgacgctcg tcgacgtggt tgggacggac
      961 ggacttgctg tcgagatact tcggacggta ggtgagactg aaagcggaca catggcctac
     1021 agcgccctac aatcggcgtt cagtcagtac gataggacca cggtctcgcg gcgattgaac
     1081 gagaagctaa tcgagtatgg tctttgtgag aagtacggcg ctcgcggaaa cctacgaatc
     1141 gagattctcg acaaaggagc gttgtacctc gaagaattag ccaaagagaa agagcgacaa
     1201 aaggcccttc aggactacgt tacgaacacc ccaaaaccaa accacaatac ccgtgtaatc
     1261 acggaccgcc ctcggggagg gagggaggac gaccaccgac cgaccgccga aggtgaggcg
     1321 tcgacgacgc cggacggacg ggttcgctta cctcggtatc acggtgtgga ctatctttcg
     1381 tacgcagaga cggtctcagc ggcgaagaca ccgccgaaga aaggcgttag tgtgacaaac
     1441 taccctgtcg aacccaaatc tgaccgggga gaagccgggt ggggctacct tgaggacgag
     1501 aatacgctcg tcgtctctgc agagtacgac tctgccatgt ctcatgtggt gatgattgca
     1561 cgagcgttac tcgatcctaa gacgttcgac caggtcctta ccgaggaccg tctcgcggaa
     1621 cttgatggac taatcgaaga tgggacgtac gttcgtgggt ctcgaaacct cgggtggttg
     1681 gctgactctg tcgactcggc gggtgagtac gtcgacgtcc tcgaggacgc gagagacgag
     1741 ctactcgaca tgacgcggtc acttgcacat gaggactacg agtgtgagac gtccgagtac
     1801 ctgagtcgaa tcacgaaaac ggcgatgggt ctcgcgggaa ccgcgtttca cgtcctcgat
     1861 ttactcgata ttgacgtcgt ttgggaggct cgcttacccg actacaaccg acacccggag
     1921 cgatacgggg aggacaatgc cgagttgtta gcaacgacgc ttgcgaagaa cgcgccgatt
     1981 gcggcgacgt acgggaacca cgtagtccgg cgactgttgt ttgaagaccg agacgagaag
     2041 cgacgtcagt cgttcgaccc tgtcgtcgac gcgagcaatc cgtatgcgaa cctgattgcg
     2101 tcgatctcgg tggttggtga cttcgggaat cgggcggacc acgtcgccgg cgtaatagaa
     2161 gaccgactgt cgaaccctgg ggagattcac gaggacgcgc cggagattgc gctcaaagtc
     2221 cctgtggtcg tcgagcatgg tccgagtacg gttgcacgag taacacgggc aatgtgtcgg
     2281 gcgaaaggtc tcgacgcgac gagagacgcg atacggctat tctctgggtt tgcacggaca
     2341 ccgtacgacg tagcccacgc aatcgggcga gggttgtctc aagaagcgac accgagagag
     2401 attcggagta gcgaagtacg gctttcactc gcgagtctcc cgtcgaagcg tctcttagag
     2461 gacgcaaccc ccacggttcg ggcgatgatt agtacgttgc tcgcgacgaa tctgagcctt
     2521 agcaagacag agttagccga gaaagcggga atctcgacgc agtcggttcg gaaccactta
     2581 ccgacacttg tagcaatggg gttagtcgac gagaccgagg gtcagtgccg actgaacctt
     2641 tcgttcacag acgagcaaga acgccacaag cggattcttc catcgttcat gctgaccgag
     2701 tcgttcactc gagacgtcct ctttgagacg ctagaagcgc ttgatgagtt gaccgaggag
     2761 gtaatcgaga tatgggtaca atgcggacct aacgggctac cggacatcga ccgtctccat
     2821 acgatgtacg actgggtaga ctgggtacgt cccttactgt cgagtctaac cgaagaacga
     2881 gacgagacgg ttcagacgct gtctattggg ccggagattg cacagcaacc cctcacggag
     2941 gcaggttcgt gattgaacta acctttggta atatgttcgt gatatgcgtt gtattcgggt
     3001 atctcgtaat ctcgtggtat ctctcgagac agtacgttca tgattagagt agggtcgacg
     3061 aactctgaac ctatgaatcg cggtcggaca acaacccccc atggatcctt gatccggtcc
     3121 cagtagagcg cgtagacgcc gagggcggta ccgttttctt gtgtgcggtt cccttgcgta
     3181 cagttcccag cgaccttccc ggctgcgacg aagggaaaac cccttacata cggtcgtcgc
     3241 actcgcgggc aatgggcttc tttgacgacc ttcaggaccg catcgagaag gtggacagtg
     3301 tcgtctcggt cggactcgac gccgacctcg accggattcc cgaacacctc cgcgacgagg
     3361 acctgccgcg gtgggcgttc aaccgccgca tcatcgacgc gacgcacgaa cacgcggcgg
     3421 cctacaagcc gaacgcggcc ttctacgagg acgaagacgg ctggcgctcg ctccgcgaga
     3481 ccatcgccta cgcacacgga aaggatgttc ccgtcctcct cgacgccaag cgcgccgaca
     3541 tcggcaacac gacgcgacag tacgcgaacc tgcttgacga cgccgatgcc atcaccgtca
     3601 acccctacat gggccgcgat tccatccagc cgttcctcgc ccgcgaggac aagggcgtgt
     3661 tcgtcctctg tcgcacctct aacccgggca gcgcggacct ccagtcgctc gaactcgcct
     3721 ccggagaacc cctgtacgag cgcgtcgcgg cgctctgcga cctctggaac gaaaacgaca
     3781 acgtcggact cgtcgtcggc gcgaccggcc ccgaagagtt ggaagacctc cgcgagcagg
     3841 tccccgacct gccgtttctc gtccccggcg tcggcgcaca gggtggcgat gccgaggcgg
     3901 cagtcgagta cggactggca gacggcgtcg gtctcgtgaa ctcctcacgc ggcatcatct
     3961 tcgccggcga agaccaggga gaacacttcg acaaggctgc gggacaggcg gcgaagcgac
     4021 tcaaagagcg gctgaaccag taccggtaag catgcaccat tccttgcggc ggcggtgctc
     4081 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
     4141 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     4201 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     4261 gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc
     4321 ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc
     4381 gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg
     4441 ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt
     4501 ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat
     4561 gacgaccatc agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc
     4621 attggaccgc tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg
     4681 gcatggattg taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca
     4741 tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca
     4801 ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt
     4861 ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca
     4921 gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag
     4981 gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa
     5041 gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc
     5101 gtgtttcgta aagtctggaa acgcggaagt cagcgctctt ccgcttcctc gctcactgac
     5161 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5221 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5281 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5341 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5401 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5461 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5521 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5581 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5641 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5701 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5761 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5821 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5881 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5941 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6001 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6061 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6361 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     6421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6661 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     6721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     7081 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     7141 caagaattct catgtttgac agcttatcat cgataagctt ctttgattcg agccatttgt
     7201 tcctcgccgg ttaggtggga gagttcgccc gattcgagta gttggttcac ctcggggtaa
     7261 ctgagttcgt ccggctcgcg ctcccgtaac tcgtcgagcg gctcgccccg ttttacatta
     7321 acctcgacct cgacctcctc gaccgaatac ccgcgctgga ggtatgcctg tctaacaccg
     7381 ctcgcggcaa tcccctcgaa agcgtccttt gccaaatcgg acggttcgac ggtgtcggtg
     7441 atcccgagat ataggagagc gaacgaccac gacaggacgt ttagcgtatg ctctttcccg
     7501 tctaaatcgc cgtcctcgaa ggcttccgtt ctaaggtcct cgtcgagttc cgaaaccagg
     7561 agcggcatat catgtagggc ttcatggagc cgctcgacta tcgctcgccg gcgttcgtat
     7621 cctgcctgtt tgctatactc cttcggtttt tggagatacc ggcggtcggc cggtgacaaa
     7681 atccctcttg ccctttcgtc aagactcatg ctttaccaat cacatcttca tgtccgtaat
     7741 gctttatacg ttgtgcccta aagcgggata tatgtctgcc acggacacgc ggattccggt
     7801 gtcgaaggat gtgcggcgcg
//
