LOCUS       sequence-343840-        7939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmids carrying a mini-CRISPR array were used to express 
            crRNAs; article demonstrates use in Haloferax mediterranei.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7939)
  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 7939)
  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-crRNA
FEATURES             Location/Qualifiers
     source          1..7939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2163..2180)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2334..2922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2414..2433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3093..3884,3885..3953))
                     /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     3716..3735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3954..4058)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4126..4144
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atccaatggt gtcgaaggga acatatatgt tactgcaggt acaacaccga gttaggaggt
       61 tacagacgaa ccctagttgg gttgaagcgt cagtcgattt atgttctcaa cgtccttaca
      121 aaggttacag acgaacccta gttgggttga agcttttttt tggtaccgtt ttcttgtgtg
      181 cggttccctt gcgtacagtt cccagcgacc ttcccggctg cgacgaaggg aaaacccctt
      241 acatacggtc gtcgcactcg cgggcaatgg gcttctttga cgaccttcag gaccgcatcg
      301 agaaggtgga cagtgtcgtc tcggtcggac tcgacgccga cctcgaccgg attcccgaac
      361 acctccgcga cgaggacctg ccgcggtggg cgttcaaccg ccgcatcatc gacgcgacgc
      421 acgaacacgc ggcggcctac aagccgaacg cggccttcta cgaggacgaa gacggctggc
      481 gctcgctccg cgagaccatc gcctacgcac acggaaagga tgttcccgtc ctcctcgacg
      541 ccaagcgcgc cgacatcggc aacacgacgc gacagtacgc gaacctgctt gacgacgccg
      601 atgccatcac cgtcaacccc tacatgggcc gcgattccat ccagccgttc ctcgcccgcg
      661 aggacaaggg cgtgttcgtc ctctgtcgca cctctaaccc gggcagcgcg gacctccagt
      721 cgctcgaact cgcctccgga gaacccctgt acgagcgcgt cgcggcgctc tgcgacctct
      781 ggaacgaaaa cgacaacgtc ggactcgtcg tcggcgcgac cggccccgaa gagttggaag
      841 acctccgcga gcaggtcccc gacctgccgt ttctcgtccc cggcgtcggc gcacagggtg
      901 gcgatgccga ggcggcagtc gagtacggac tggcagacgg cgtcggtctc gtgaactcct
      961 cacgcggcat catcttcgcc ggcgaagacc agggagaaca cttcgacaag gctgcgggac
     1021 aggcggcgaa gcgactcaaa gagcggctga accagtaccg gtaagcatgc accattcctt
     1081 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     1141 cataagggag agcgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     1201 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     1261 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     1321 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     1381 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     1441 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc
     1501 cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg
     1561 tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc
     1621 agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc
     1681 acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg
     1741 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg
     1801 ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg
     1861 cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc
     1921 ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt
     1981 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
     2041 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     2101 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ctcttccgct
     2161 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2221 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2281 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2341 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2401 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2461 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2521 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2581 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2641 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2701 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2761 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2821 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2881 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2941 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3001 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3061 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3121 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3181 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3241 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3301 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3361 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     3421 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3481 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3541 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3601 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3661 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     3721 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3781 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3841 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3901 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3961 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4021 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4081 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4141 aggccctttc gtcttcaaga attctcatgt ttgacagctt atcatcgata agcttctttg
     4201 attcgagcca tttgttcctc gccggttagg tgggagagtt cgcccgattc gagtagttgg
     4261 ttcacctcgg ggtaactgag ttcgtccggc tcgcgctccc gtaactcgtc gagcggctcg
     4321 ccccgtttta cattaacctc gacctcgacc tcctcgaccg aatacccgcg ctggaggtat
     4381 gcctgtctaa caccgctcgc ggcaatcccc tcgaaagcgt cctttgccaa atcggacggt
     4441 tcgacggtgt cggtgatccc gagatatagg agagcgaacg accacgacag gacgtttagc
     4501 gtatgctctt tcccgtctaa atcgccgtcc tcgaaggctt ccgttctaag gtcctcgtcg
     4561 agttccgaaa ccaggagcgg catatcatgt agggcttcat ggagccgctc gactatcgct
     4621 cgccggcgtt cgtatcctgc ctgtttgcta tactccttcg gtttttggag ataccggcgg
     4681 tcggccggtg acaaaatccc tcttgccctt tcgtcaagac tcatgcttta ccaatcacat
     4741 cttcatgtcc gtaatgcttt atacgttgtg ccctaaagcg ggatatatgt ctgccacgga
     4801 cacgcggatt ccggtgtcga aggatgtgcg gcgcgacctc cgggttctca aggcgcggga
     4861 aggccgacgg tcatacgatg agacaattgc ggtggtgctt gacgcatatc tctctgagaa
     4921 ggtggattaa taaactatag tcgtgactag cgaactagtt gaatcaaacg acgccgggag
     4981 cacacgagtc tgccaggact tcgctcctat cggcgtcgaa ggatgcttac gcggacacgg
     5041 cgggcgggtg gcccgttgtc ctaactcaac caatgtggtt ggggcttatg ggattatcgg
     5101 attttcaccc ggcgttagtg ttcgcacaaa agcgacacta aatgtacgat acggacgata
     5161 cgacggcaca cgaaccgact gagaagcgaa gccttcgctt tgcgactgac ctcgaggcgg
     5221 acgtctcgct tacagactgg accgcccttc gggttcatcg agtaatcttc gacgagacag
     5281 agagtcaatt tgcgcttctc gaggcactac aaacagcggc ggacggtgtc gacatactcg
     5341 agtttgaaca ccgactgtcg agttgtgggt ttgacgtcga tgggtcagtc gtgaaacttg
     5401 cacgacgagc tgtcgccggc gagagacggg ctctgtggga cttgattcgg tatgcgcgac
     5461 ggttgggaca tgatgagatg ggacagccac agagcgcctt agaggacgct ctcgcgttca
     5521 ctcgtctatt gggctatagc aaccctcaac agacgatagc aatcggtctc aatgcggctt
     5581 tcagagagac gcggcgcgac caggccgacg agattatcga gtacgttcta acgctgtctg
     5641 agagcgtcga cgtggtcctt ggtgggtcag agatcgaccg agtgtacttg cacaactact
     5701 acggcgatat tctcccatcg agtgttacga atccgtgcaa cgacacccgt aatgattact
     5761 ccattacgac gctcgtcgac gtggttggga cggacggact tgctgtcgag atacttcgga
     5821 cggtaggtga gactgaaagc ggacacatgg cctacagcgc cctacaatcg gcgttcagtc
     5881 agtacgatag gaccacggtc tcgcggcgat tgaacgagaa gctaatcgag tatggtcttt
     5941 gtgagaagta cggcgctcgc ggaaacctac gaatcgagat tctcgacaaa ggagcgttgt
     6001 acctcgaaga attagccaaa gagaaagagc gacaaaaggc ccttcaggac tacgttacga
     6061 acaccccaaa accaaaccac aatacccgtg taatcacgga ccgccctcgg ggagggaggg
     6121 aggacgacca ccgaccgacc gccgaaggtg aggcgtcgac gacgccggac ggacgggttc
     6181 gcttacctcg gtatcacggt gtggactatc tttcgtacgc agagacggtc tcagcggcga
     6241 agacaccgcc gaagaaaggc gttagtgtga caaactaccc tgtcgaaccc aaatctgacc
     6301 ggggagaagc cgggtggggc taccttgagg acgagaatac gctcgtcgtc tctgcagagt
     6361 acgactctgc catgtctcat gtggtgatga ttgcacgagc gttactcgat cctaagacgt
     6421 tcgaccaggt ccttaccgag gaccgtctcg cggaacttga tggactaatc gaagatggga
     6481 cgtacgttcg tgggtctcga aacctcgggt ggttggctga ctctgtcgac tcggcgggtg
     6541 agtacgtcga cgtcctcgag gacgcgagag acgagctact cgacatgacg cggtcacttg
     6601 cacatgagga ctacgagtgt gagacgtccg agtacctgag tcgaatcacg aaaacggcga
     6661 tgggtctcgc gggaaccgcg tttcacgtcc tcgatttact cgatattgac gtcgtttggg
     6721 aggctcgctt acccgactac aaccgacacc cggagcgata cggggaggac aatgccgagt
     6781 tgttagcaac gacgcttgcg aagaacgcgc cgattgcggc gacgtacggg aaccacgtag
     6841 tccggcgact gttgtttgaa gaccgagacg agaagcgacg tcagtcgttc gaccctgtcg
     6901 tcgacgcgag caatccgtat gcgaacctga ttgcgtcgat ctcggtggtt ggtgacttcg
     6961 ggaatcgggc ggaccacgtc gccggcgtaa tagaagaccg actgtcgaac cctggggaga
     7021 ttcacgagga cgcgccggag attgcgctca aagtccctgt ggtcgtcgag catggtccga
     7081 gtacggttgc acgagtaaca cgggcaatgt gtcgggcgaa aggtctcgac gcgacgagag
     7141 acgcgatacg gctattctct gggtttgcac ggacaccgta cgacgtagcc cacgcaatcg
     7201 ggcgagggtt gtctcaagaa gcgacaccga gagagattcg gagtagcgaa gtacggcttt
     7261 cactcgcgag tctcccgtcg aagcgtctct tagaggacgc aacccccacg gttcgggcga
     7321 tgattagtac gttgctcgcg acgaatctga gccttagcaa gacagagtta gccgagaaag
     7381 cgggaatctc gacgcagtcg gttcggaacc acttaccgac acttgtagca atggggttag
     7441 tcgacgagac cgagggtcag tgccgactga acctttcgtt cacagacgag caagaacgcc
     7501 acaagcggat tcttccatcg ttcatgctga ccgagtcgtt cactcgagac gtcctctttg
     7561 agacgctaga agcgcttgat gagttgaccg aggaggtaat cgagatatgg gtacaatgcg
     7621 gacctaacgg gctaccggac atcgaccgtc tccatacgat gtacgactgg gtagactggg
     7681 tacgtccctt actgtcgagt ctaaccgaag aacgagacga gacggttcag acgctgtcta
     7741 ttgggccgga gattgcacag caacccctca cggaggcagg ttcgtgattg aactaacctt
     7801 tggtaatatg ttcgtgatat gcgttgtatt cgggtatctc gtaatctcgt ggtatctctc
     7861 gagacagtac gttcatgatt agagtagggt cgacgaactc tgaacctatg aatcgcggtc
     7921 ggacaacaac cccccatgg
//
