LOCUS       sequence-469952-        9746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid pQdCas12a.luxR(mut)-sggfp(B) expresses the dCas12a 
            endonuclease and the sgRNA (design B) targeting the gfp gene. 
            Additionally, it contains a mutation in the luxR gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9746)
  AUTHORS   Golla SA, Abo-Hashesh M, Gupta D, Liu Y, Mahadevan R
  TITLE     Model-Based Optimization of a qCRISPRi Circuit for Dynamic Control 
            of Metabolic Pathways.
  JOURNAL   ACS Synth Biol. 2025 Oct 17;14(10):3890-3898. doi: 
            10.1021/acssynbio.5c00095. Epub 2025 Sep 29.
  PUBMED    41021780
REFERENCE   2  (bases 1 to 9746)
  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: 
            "10.1021/acssynbio.5c00095"; journalName: "ACS Synth Biol"; date: 
            "2025-10-17- 17"; volume: "14"; issue: "10"; pages: "3890-3898"
COMMENT     Sequence Label: pQdCas12a.luxR(mut)-sggfp(B)
FEATURES             Location/Qualifiers
     source          1..9746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             514..525
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(2262..3056)
                     /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
                     SHLAPVEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2373..2392)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2983..3002
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4536..5307
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5308..5970
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGE*QLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     promoter        complement(6310..6344)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 gattatttct aacgcctcat ctatatctgt gatatcacta ttggctttaa atagttctat
       61 accattatcc ttagattgct ttagccatag aattaaatct gactcttgcc cttttttagc
      121 atcgataagg ttttgattaa acaaattctt aaatttctct gagtccttta tatattcaga
      181 tatttgtttc tttatcgtat cttttgcact tttaaaatct ttttgtagat tatcatcatc
      241 actcttttta agtttaaaat aaacatcaga atagttttgt aataaatctt cgctaataca
      301 aaccgaactt aatatctcct ctataaaaaa ctgatgatat ttatcaatta tttgtttagc
      361 ctttttgtag tctttagctc ttttctcatc atctaaaatc aaacctcttg cttttatgtt
      421 ttcaagtgtt ttaccctgtg ggattaactc aaatcttaga gttttactta aactatattt
      481 attaacaaat tcttgataaa ttgacataga tcctttctcc tcttttttat tcgactataa
      541 caaaccattt tcttgcgtaa acctgtacga tcctacaggt agatctttaa tttttaaaaa
      601 atggccaatc aattgctcct gttaaaattg ctttagaaat actttggcag cggtttgttg
      661 tatcgagttt catttgcgca ttggttaaat ggaaagtgac agtacgctca ctgcagccta
      721 atatttttga aatatcccaa gagctttttc cttcgcatgc ccacgctaaa cattcttttt
      781 ctcttttggt taaatcgttg tttgatttat tatttgctat atttattttt cgataattat
      841 caactagaga aggaacaatt aatggtatgt tcatacacgc atgtaaaaat aaactatcta
      901 tatagttgtc aatttctgaa tgtgcaaaac taagcattcc gaagccattg ttagccgtat
      961 gaatagggaa actaaaccca gtgataagac ctgatgtttt cgcttcttta attacatttg
     1021 gagatttttt atttacagca ttgttttcaa atatattcca attaattggt gaatgattgg
     1081 atttagaata atctactata ggatcatatt ttattaaatt agcgcgccat aatattgcct
     1141 ccatttttta gggtaattat ctagaattga aatatcagat ttaaccatag aatgaggata
     1201 aatgatcgcg agtaaataat attcacaatg taccatttta gtcatatcag ataagcattg
     1261 attaatatca ttattgctgc cacaagcttt aattttatta attattctgt atgtgtcgtc
     1321 ggcatttatg tttttcatac ccatctcttt atccttacct attgtttgtc gcaagttttg
     1381 cgtgttatat atcattaaaa cggtaatgga ttgacatttg attctaataa attggatttt
     1441 tgtcacacta ttgtatcgct gggaatacaa ttacttaaca taagcacctg taggatcgta
     1501 caggtttacg caagaaaatg gtttgttata gtcgaataaa cgcaagggag gttggtatga
     1561 ctataatgat aaaaaaatcg gattttttgg caattccatc ggaggagtat aaaggtattc
     1621 taagtcttcg ttatcaagtg tttaagcaaa gacttgagtg ggacttagtt gtagaaaata
     1681 accttgaatc agatgagtat gataactcaa atgcagaata tatttatgct tgtgatgata
     1741 ctgaaaatgt aagtggatgc tggcgtttat tacctacaac aggtgattat atgctgaaaa
     1801 gtgtttttcc tgaattgctt ggtcaacaga gtgctcccaa agatcctaat atagtcgaat
     1861 taagtcgttt tgctgtaggt aaaaatagct caaagataaa taactctgct agtgaaatta
     1921 caatgaaact atttgaagct atatataaac acgctgttag tcaaggtatt acagaatatg
     1981 taacagtaac atcaacagca atagagcgat ttttaaagcg tattaaagtt ccttgtcatc
     2041 gtattggaga caaagaaatt catgtattag gtgatactaa atcggttgta ttgtctatgc
     2101 ctattaatga acagtttaaa aaagcagtct taaattaatc tagataagtt caggatgaat
     2161 tctctagtcc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     2221 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     2281 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     2341 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     2401 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     2461 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     2521 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     2581 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     2641 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     2701 gcatcagcca tgatggatac tttctcgaca ggagcaaggt gagatgacag gagatcctgc
     2761 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     2821 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgtagt
     2881 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     2941 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     3001 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     3061 aacgatcctc atcctgtctc ttgatcaaat cttgatcccc tgcgccatca gatccttggc
     3121 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     3181 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta
     3241 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     3301 cagtagctga cattcatccc aggtggcact tttcggggaa atgtgcgcgc ccgcgttcct
     3361 actggcgctg ggcctgtttc tggcgctgga cttcccgctg ttccgtcagc agcttttcgc
     3421 ccacggcctt gatgatcgcg gcggccttgg cctgcatatc ccgattcaac ggccccaggg
     3481 cgtccagaac gggcttcagg cgctcccgaa ggtctcgggc cgtctcttgg gcttgatcgg
     3541 ccttcttgcg catctcacgc gctcctgcgg cggcctgtag ggcaggctca tacccctgcc
     3601 gaaccgcttt tgtcagccgg tcggccacgg cttccggcgt ctcaacgcgc tttgagattc
     3661 ccagcttttc ggccaatccc tgcggtgcat aggcgcgtgg ctcgaccgct tgcgggctga
     3721 tggtgacgtg gcccactggt ggccgctcca gggcctcgta gaacgcctga atgcgcgtgt
     3781 gacgtgcctt gctgccctcg atgccccgtt gcagccctag atcggccaca gcagccgcaa
     3841 acgtggtctg gtcgcgggtc atctgcgctt tgttgccgat gaactccttg gccgacagcc
     3901 tgccgtcctg cgtcagcggc accacgaacg cggtcatgtg cgggctggtt tcgtcacggt
     3961 ggatgctggc cgtcacgatg cgatccgccc cgtacttgtc cgccagccac ttgtgcgcct
     4021 tctcgaagaa cgccgcctgc tgttcttggc tggccgactt ccaccattcc gggctggccg
     4081 tcatgacgta ctcgaccgcc aacacagcgt ccttgcgccg cttctctggc agcaactcgc
     4141 gcagtcggcc catcgcttca tcggtgctgc tggccgccca gtgctcgttc tctggcgtcc
     4201 tgctggcgtc agcgttgggc gtctcgcgct cgcggtaggc gtgcttgaga ctggccgcca
     4261 cgttgcccat tttcgccagc ttcttgcatc gcatgatcgc gtatgccgcc atgcctgccc
     4321 ctcccttttg gtgtccaacc ggctcgacgg gggcagcgca aggcggtgcc tccggcgggc
     4381 cactcaatgc ttgagtatac tcactagact ttgcttcgca aagtcgtgac cgcctacggc
     4441 ggctgcggcg ccctacgggc ttgctctccg ggcttcgccc tgcgcggtcg ctgcgctccc
     4501 ttgccagccc gtggatatgt ggacgatggc cgcgagcggc caccggctgg ctcgcttcgc
     4561 tcggcccgtg gacaaccctg ctggacaagc tgatggacag gctgcgcctg cccacgagct
     4621 tgaccacagg gattgcccac cggctaccca gccttcgacc acatacccac cggctccaac
     4681 tgcgcggcct gcggccttgc cccatcaatt tttttaattt tctctgggga aaagcctccg
     4741 gcctgcggcc tgcgcgcttc gcttgccggt tggacaccaa gtggaaggcg ggtcaaggct
     4801 cgcgcagcga ccgcgcagcg gcttggcctt gacgcgcctg gaacgaccca agcctatgcg
     4861 agtgggggca gtcgaagggc gaagcccgcc cgcctgcccc ccgagcctca cggcggcgag
     4921 tgcgggggtt ccaagggggc agcgccacct tgggcaaggc cgaaggccgc gcagtcgatc
     4981 aacaagcccc ggaggggcca ctttttgccg gagggggagc cgcgccgaag gcgtggggga
     5041 accccgcagg ggtgcccttc tttgggcacc aaagaactag atatagggcg aaatgcgaaa
     5101 gacttaaaaa tcaacaactt aaaaaagggg ggtacgcaac agctcattgc ggcacccccc
     5161 gcaatagctc attgcgtagg ttaaagaaaa tctgtaattg actgccactt ttacgcaacg
     5221 cataattgtt gtcgcgctgc caaaaagttg cagctgattg tgcatggtgc cgcaaccgtg
     5281 cggcacccct accgcatgga gataagcatg gccacgcagt ccagagaaat cggcattcaa
     5341 gccaagaaca agcccggtca ctgggtgcaa acggaacgca aagcgcatga ggcgtgggcc
     5401 gggcttattg cgaggaaacc cacggcggca atgctgctgc atcacctcgt ggcgcagatg
     5461 ggccaccaga acgccgtggt ggtcagccag aagacacttt ccaagctcat cggacgttct
     5521 ttgcggacgg tccaatacgc agtcaaggac ttggtggccg agcgctggat ctccgtcgtg
     5581 aagctcaacg gccccggcac cgtgtcggcc tacgtggtca atgaccgcgt ggcgtggggc
     5641 cagccccgcg accagttgcg cctgtcggtg ttcagtgccg ccgtggtggt tgatcacgac
     5701 gaccaggacg aatcgctgtt ggggcatggc gacctgcgcc gcatcccgac cctgtatccg
     5761 ggcgagtagc aactaccgac cggccccggc gaggagccgc ccagccagcc cggcattccg
     5821 ggcatggaac cagacctgcc agccttgacc gaaacggagg aatgggaacg gcgcgggcag
     5881 cagcgcctgc cgatgcccga tgagccgtgt tttctggacg atggcgagcc gttggagccg
     5941 ccgacacggg tcacgctgcc gcgccggtag cacttgggtt gcgcagcaac ccgtaagtgc
     6001 gctgttccag actatactag actgcaggtc ctgaagttaa caaaaaaaat gaccttcata
     6061 aatcgctaat ctacaacagt agaaattatt taaagttctt agaccggcta ctaatacaaa
     6121 aggctctaga catctacaac agtagaaatt atttaaagtt cttagactga attatctgaa
     6181 ggcacaggaa tagtagcatc tacaacagta gaaattattt aaagttctta gacctcacca
     6241 tgctcgagcc ctgatctaca acagtagaaa ttatttaaag ttcttagacc cgtttttgcc
     6301 taaatcagcg ctagcattat acctaggact gagctagctg tcaactcgag ttagttattc
     6361 ctattctgca cgaactcaaa atactcttca tttttgataa ccaaattgag ttttttgccc
     6421 tcttgattat ttttgatcct acctagtagc atcagacctt ttagcccaat atgataagca
     6481 ccattggcat cagcatcttg aggcatattt tttggcgcct gtcgcgaatc aaagaaattg
     6541 ccatttacat ctgctactgg tgaaattaga taatctaact cagtacctgt ttttgagtta
     6601 cgcatttgta agatagtatt taggacacta gttagcttag caaaaaactt tttgtcgctc
     6661 tcaccgcaaa tagctgcttt gatacattcg ccatgcccat attcgataga ataatctttt
     6721 agcaatttct ccaactcttt agttggataa acttctcgag tatcccaatt atgattttta
     6781 tctgaatttc taaagttaat caatctactc ccaaagctag ctatagtcca cttgcctttg
     6841 gcagccttgt caccaaagtt tttataatca aaactaaact caaaatagcc cttatcaagg
     6901 ttataacaaa tcttgtcaaa cttactaaag aactcttgag atttgctgac actttcatac
     6961 ttaggatata actgatttac aaaaccagtt acaggacaaa tttttgaagt aaaaccagct
     7021 ggtacatagt agataatacc tgtttgttta cccatctttt taaaagtctc aaaaggtgct
     7081 gttagctgat aagctctaag cactccccca gttttatcaa actcattatc tttgaaaact
     7141 agatagttta gtttctcaat tagcattttt tctaactttt gatagacctg cttctctacc
     7201 ttgaaacgcc ctcttttaaa tccaaaattt aaatcctcaa aaaccacaat agcattatac
     7261 tctataacta gcttagctat ttcatgaact acctgagata gatagccctc tttcatctct
     7321 ttgatgttat ttatcttttt ccagtctttc ctagctgaat ccctatcttt ctctattgca
     7381 gcaagcttat catggtagtt tgttttcatt ctatcattac caatgatgtt gaaagtatct
     7441 tgtttgatga tattgccttt accatctacc aaagtatagt aagctaaatg tctttcacct
     7501 ctagctatac ttaatatatg aacatcattt gctttttctt ttagcaataa attgatttca
     7561 tcattaaact tattagctcc actagattta aaattgattg taataggaca gtgaaagaaa
     7621 aacttatctt cagtaaagcg tttatctttg attaaatcat attcaaaaac actctctttt
     7681 ttaggattat ctttgttttt attagctatt gcctctttag ctgggtgagt gattttttta
     7741 ggtattgatt gtttacgata aaaaagctct gcctcaccat ttagcttata aaccacatct
     7801 tgaagatttc tctcatcaaa cagcgctttc caatataaag tatgtagatt tggtcgccct
     7861 ttgctataag ctgaaaaatc tttattatag atttggaata ggtacaattt accctgatta
     7921 actacgctat caatatagct ctctgatata ttttcaaaag ttagtttgta gccttgattt
     7981 tcaacttctc tataaaattc atctatagaa ttatatcttt gagtatcaga aaatctaaat
     8041 ccaaaatctt tccactccgg atgcttactt atagactgtt tataaaaatc tataaatttt
     8101 cggcaatctt caatattaaa ctcaaatttt tcatatcctt tttgaggact accatttttt
     8161 gtatgtgtgg aatgatttct tattctaagt atatcttcac taggattata aaattttata
     8221 gatttagcag aaaagaaaac cttaggtaac attttatttg cgccaggtaa aagtttataa
     8281 acaatttttt tataaccctc gcctttattt tctttgatag ctttatcatc aaatattttg
     8341 ttatttttct tattcatcac acccagataa tatttatcat ctttgataaa taaaattgcc
     8401 gtattgtcag gctctttatt tttatcccaa ccattagcca aagtcgagtt ctcaaaattg
     8461 agcttaaatt tctcatcact atatggcttt tgagttatat agtttctaat tttgttataa
     8521 agaggcacta tattcgctag ctcaaagtag cactcctcaa atactagata aaaatgctca
     8581 tccttgtcta aaatatttgc cttatcttct gactgactaa tatgaaatat ttttagttta
     8641 tgtaagagat tattagtttg atctaaaaga tccttgatag ctttaacatc atcttccgca
     8701 ctagcttgaa gtaggtcttt tttaccttga ttttgatatt tgatagatat ctgtgccaaa
     8761 ttgtctttgt tttgagctat ttcatcaaat atcatcggaa tagccgcaaa gtttgcaagt
     8821 atttcttcaa acctacactg tttatctata tctctatgct tattaaattc ttctaaggca
     8881 agctttatag tttctagaga taagtatttt gctttttcag tttttttggc tattaattct
     8941 tgctctttct tactagggtt atcaagattt ttaggtgcta tttgttgagt tatatattct
     9001 agtaccgctg taccaataac actataatca tcaaaaactt gttgtgatag atcagtaaga
     9061 gatttatcat ttttaaaata aattttactc aaatcaagtt tttgagcttt taaatcatca
     9121 aataataaag atagtgtttc tttaatagat ttttcttcta ctgttttaaa agctgctatt
     9181 tgctcataaa aactttgcat cgttgtaact acatcactat catcttctaa cttatcaatt
     9241 acaaaagatt tagattctgt atcacttaaa atttgcttaa ataaaacact cattttatat
     9301 tttttgagtg ttttatcatt tatttgctgt gagtatagat ttatatattc atttatacct
     9361 tttctctttg tattttcacc atttacaaat ttaccaccaa taatagtatt aaatttagta
     9421 ataccacttt gatttagata attattaaag tttgctatct caaaaacttc atcaagtgaa
     9481 aaaactcttt gattaacttc agatgttttg tagtcaatat caaaggttag ctcttctgcc
     9541 aaatcttttt taatttgttc atagtttata gcttctggag ctttgtcttt taaactctca
     9601 tacttagctt tattttctag aaatttaggc aaattatcat ctactatcct ataaataata
     9661 gatgtaggaa tatcattgct actataaaca ttttttctat tttcatgaaa acccttaaaa
     9721 taagttgtcc aacctttaaa agattt
//
