LOCUS       sequence-344139-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBM23 contained pyrF and mini-CRISPR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5987)
  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 5987)
  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: pBM23-pyrF-CS
FEATURES             Location/Qualifiers
     source          1..5987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1865..1883)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1866..1883)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1904..1926)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2120..2139)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2239..2261
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2299..2317)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2385..2489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2490..2558,2559..3350)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGAQVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEEDKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2708..2727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3664..4252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4153..4172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4436..5230)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4436..5230)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4547..4566)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5157..5176
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5426..5443
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5560..5581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5596..5601,5602..5619,5620..5626)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5634..5650
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5639..5661
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5683..5702
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5683..5701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgttcccggt atttgccgtc gttcgcggcg taaacaacgt ctcgggcgtc ctcggaaacc
       61 agtacatcaa cgaccgtgtc gactcggttg ggcgcgcaac cgtcctctcc gccgccggga
      121 tgctctactc gctcgccgtc gttccgttcg aactcgcggg tggtgtcgtc gccgacgtgg
      181 tctctccaac cggcgcactc gccctattcg gcggggttct tgcggtcggt gcaggcgtgt
      241 tgtgggttgt cgaacgccca atctgaaatg gtgtcgaagg gaacatatat gttactgcag
      301 gtacaacacc gagttaggag gttacagacg aaccctagtt gggttgaagc atatggtaag
      361 caaccctttg ggtatgtcag gcgaagttac agacgaaccc tagttgggtt gaagcgtgtt
      421 ctcagaaaga atatggttcg aggccatcta gttacagacg aaccctagtt gggttgaagc
      481 ttttttttgt tttcttgtgt gcggttccct tgcgtacagt tcccagcgac cttcccggct
      541 gcgacgaagg gaaaacccct tacatacggt cgtcgcactc gcgggcaatg ggcttctttg
      601 acgaccttca ggaccgcatc gagaaggtgg acagtgtcgt ctcggtcgga ctcgacgccg
      661 acctcgaccg gattcccgaa cacctccgcg acgaggacct gccgcggtgg gcgttcaacc
      721 gccgcatcat cgacgcgacg cacgaacacg cggcggccta caagccgaac gcggccttct
      781 acgaggacga agacggctgg cgctcgctcc gcgagaccat cgcctacgca cacggaaagg
      841 atgttcccgt cctcctcgac gccaagcgcg ccgacatcgg caacacgacg cgacagtacg
      901 cgaacctgct tgacgacgcc gatgccatca ccgtcaaccc ctacatgggc cgcgattcca
      961 tccagccgtt cctcgcccgc gaggacaagg gcgtgttcgt cctctgtcgc acctctaacc
     1021 cgggcagcgc ggacctccag tcgctcgaac tcgcctccgg agaacccctg tacgagcgcg
     1081 tcgcggcgct ctgcgacctc tggaacgaaa acgacaacgt cggactcgtc gtcggcgcga
     1141 ccggccccga agagttggaa gacctccgcg agcaggtccc cgacctgccg tttctcgtcc
     1201 ccggcgtcgg cgcacagggt ggcgatgccg aggcggcagt cgagtacgga ctggcagacg
     1261 gcgtcggtct cgtgaactcc tcacgcggca tcatcttcgc cggcgaagac cagggagaac
     1321 acttcgacaa ggctgcggga caggcggcga agcgactcaa agagcggctg aaccagtacc
     1381 ggtaagtctg cggaaatcac agactggcac tggactgtac cgctctcggc gggacgttta
     1441 ccagcgtgcg tcaccaacta cgtccgtgga ccgagaccga cttatcatgg gcctcgcggc
     1501 ggtgttcgcg ggtattacca tgctcctcgt cgtgttggca ttcgcctacc gacagctgtt
     1561 gttgttgttc gtagctgtgc ccttcggcgc gacgacgtac ttcatgtact cggacgtgag
     1621 cggccggttc gaagaacggt ttagacggcg agttcaggcg gatgctggcg gggtcggctt
     1681 tcagtcgggc ggtggcggcc cacagcagcg cgaaaccggc gggttcggtg cagggccacg
     1741 acgtggtccc cgagaaggcc ccggtgcagg acggttcggg cgcgccggac gacagggtgg
     1801 accgcaaagt ggcgcacgta ctgaaagggc gacacgcccg ggtaccatgg atccgagctc
     1861 gtattctata gtgtcaccta aatagcttca attggctggc cgtcgtttta caacgtcgtg
     1921 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1981 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     2041 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2101 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     2161 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2221 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2281 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     2341 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2401 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2461 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgcgctta
     2521 tcccattttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2581 taaaagatgc tgaagatcag ttgggtgcac aagtgggtta catcgaactg gatctcaaca
     2641 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2701 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2761 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2821 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2881 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2941 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3001 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3061 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3121 aggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3181 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3241 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3301 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3361 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggctct
     3421 aggtgaagag cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3481 actgagcgtc agatcccgta gaaaagatca aaccttcttc ttgagatcgt ttttttctgc
     3541 gccccgtaga aaagatcaaa gcatcttctt gagaacgatt ttttctgcgc ccgtagaaaa
     3601 gatcaaacgt tcttcttgag atgcattttt tctgcgcccc gtagaaaaga tcaaaggttc
     3661 ttcttgagat cgattttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3721 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3781 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3841 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3901 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3961 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4021 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4081 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4141 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4201 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4261 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4321 tgcgttatcc cctgattctg tggataaccg tattaccgcc agatccgcgg tgtgaaatac
     4381 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta ataattcaga
     4441 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     4501 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     4561 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     4621 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     4681 gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     4741 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     4801 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     4861 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     4921 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     4981 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     5041 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     5101 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     5161 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     5221 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agagcttgat cccctgcgcc
     5281 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     5341 cagagggcgc cccagctggc aattccggtt cgcttgctgt ccagagtgag ctgataccgc
     5401 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5461 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5521 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5581 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5641 cggataacaa tttcacacag gaaacagcta tggatggaat tgtaatacga ctcactatag
     5701 ggcaaagctt gcatgcctcg agctgcagtc tagacccggg cgtgtcgccc ttgcgtacat
     5761 cctcgaccag ccgatactcc gtgctgtcgc agtggacctt ggtatccccg agtcagcgac
     5821 aaatacggcc gtgagtacct cgtatgccgc cttctcgatt atcgcggcgg gggcaaccta
     5881 ccgttccggg tggattagcg accgactcgg cgtccgacgg tggttcctcg tctcgccgct
     5941 tctcgtcgcc gtgtcctacg ccgtgctccc aatcttcggt ccactgg
//
