LOCUS       sequence-344135-        4638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids for gene knockout were constructed based on suicide plasmid
            pHFX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4638)
  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 4638)
  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: pHFX-cas3UD
FEATURES             Location/Qualifiers
     source          1..4638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      169..757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     658..677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     911..928
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1045..1066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1081..1086,1087..1104,1105..1111)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1119..1135
                     /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     1124..1146
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1143..1159
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1143..1159
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1183..1202
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1183..1201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3000..3018)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3032..3049)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3032..3048)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3041..3063)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3190..3645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3277..3296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3487..3508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3671..3775
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3776..3844,3845..4636)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3994..4013)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
       61 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
      121 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      181 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      241 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      301 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      361 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      421 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      481 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      541 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      601 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      661 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      721 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      781 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      841 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      901 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      961 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     1021 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     1081 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     1141 cacaggaaac agctatgacc atgattacgc cagatatcaa attaatacga ctcactatag
     1201 ggagaagctt gcatgccacg cgtctcgagg gcccctgcag gtcgactcta gaggatccgc
     1261 tcgttgaccc ggcgtaccgt ctctatgtct ggatgagtga ttcccattgg ttcgagacgc
     1321 tccacgagac gctcgacgaa ggcaaatcac actacgttcc gagtcttggt ctttctgagt
     1381 acctcgcaga gatcacgtac cacggacgct tcgaggttga gtctggaccg actgacactg
     1441 cggtggcagt cgactctgcc gtcccgaacg cggtcgatca tgtcgttcca gacgctgaat
     1501 cccggtgtca aatcgaagag tcgcccgcgt tcatgacagt tgacggaggc ggacgaacaa
     1561 cgactgactt caccagctac acgtacaatc cagacgcagg accggttcgg gtacgaaatc
     1621 cggacacggc tatcgtcgac ggaaacacgg tgatgttcgt ttgagatgac cgacgtcgat
     1681 ccagttcaac gactcctgcg aactgcgaga gatgacgcgc gagacgaatc gtttcgggta
     1741 accggcgtca tgatgcagta ttacgtcgtc tgtaagcgcg aactctggtt tcacagccgt
     1801 catatcgaga ttgaccgcgg gaactccgct attgtccgag gaacccatat cgacgaaacc
     1861 gcgtatagcg acaaacggcg acatgtctcg attgacagta caattgcgat tgacgtattg
     1921 gatgatggtc gtgtgatgga ggtcaaacct tcctcggcac tcgtcgaacc tgcgaaactc
     1981 cagttactct actacttatg gtatctgaag cacgttgttg gtgtcgagaa atctggcgta
     2041 cttgcacacc cgacagagcg caagcgagaa gacgtagaat tgactgggta ccttaccggt
     2101 actggttcag ccgctctttg agtcgcttcg ccgcctgtcc cgcagccttg tcgaagtgtt
     2161 ctccctggtc ttcgccggcg aagatgatgc cgcgtgagga gttcacgaga ccgacgccgt
     2221 ctgccagtcc gtactcgact gccgcctcgg catcgccacc ctgtgcgccg acgccgggga
     2281 cgagaaacgg caggtcgggg acctgctcgc ggaggtcttc caactcttcg gggccggtcg
     2341 cgccgacgac gagtccgacg ttgtcgtttt cgttccagag gtcgcagagc gccgcgacgc
     2401 gctcgtacag gggttctccg gaggcgagtt cgagcgactg gaggtccgcg ctgcccgggt
     2461 tagaggtgcg acagaggacg aacacgccct tgtcctcgcg ggcgaggaac ggctggatgg
     2521 aatcgcggcc catgtagggg ttgacggtga tggcatcggc gtcgtcaagc aggttcgcgt
     2581 actgtcgcgt cgtgttgccg atgtcggcgc gcttggcgtc gaggaggacg ggaacatcct
     2641 ttccgtgtgc gtaggcgatg gtctcgcgga gcgagcgcca gccgtcttcg tcctcgtaga
     2701 aggccgcgtt cggcttgtag gccgccgcgt gttcgtgcgt cgcgtcgatg atgcggcggt
     2761 tgaacgccca ccgcggcagg tcctcgtcgc ggaggtgttc gggaatccgg tcgaggtcgg
     2821 cgtcgagtcc gaccgagacg acactgtcca ccttctcgat gcggtcctga aggtcgtcaa
     2881 agaagcccat tgcccgcgag tgcgacgacc gtatgtaagg ggttttccct tcgtcgcagc
     2941 cgggaaggtc gctgggaact gtacgcaagg gaaccgcaca caagaaaacg aattctctcc
     3001 ctttagtgag ggttaattac aggcctaatt cactggccgt cgttttacaa cgtcgtgact
     3061 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3121 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3181 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3541 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3601 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     3661 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3721 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3781 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3841 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3901 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3961 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4021 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4081 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4141 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4201 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4261 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4321 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4381 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4441 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4501 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4561 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4621 gattaagcat tggtaact
//
