LOCUS       sequence-469911-        9760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid pQdCas12a.luxR(mut)-sggfp(A) expresses the dCas12a 
            endonuclease and the sgRNA (design A) 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 9760)
  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 9760)
  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(A)
FEATURES             Location/Qualifiers
     source          1..9760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             2622..2633
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(4370..5164)
                     /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(4481..4500)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5091..5110
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      6644..7415
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             7416..8078
                     /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(8432..8466)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 cgtttatctt tgattaaatc atattcaaaa acactctctt ttttaggatt atctttgttt
       61 ttattagcta ttgcctcttt agctgggtga gtgatttttt taggtattga ttgtttacga
      121 taaaaaagct ctgcctcacc atttagctta taaaccacat cttgaagatt tctctcatca
      181 aacagcgctt tccaatataa agtatgtaga tttggtcgcc ctttgctata agctgaaaaa
      241 tctttattat agatttggaa taggtacaat ttaccctgat taactacgct atcaatatag
      301 ctctctgata tattttcaaa agttagtttg tagccttgat tttcaacttc tctataaaat
      361 tcatctatag aattatatct ttgagtatca gaaaatctaa atccaaaatc tttccactcc
      421 ggatgcttac ttatagactg tttataaaaa tctataaatt ttcggcaatc ttcaatatta
      481 aactcaaatt tttcatatcc tttttgagga ctaccatttt ttgtatgtgt ggaatgattt
      541 cttattctaa gtatatcttc actaggatta taaaatttta tagatttagc agaaaagaaa
      601 accttaggta acattttatt tgcgccaggt aaaagtttat aaacaatttt tttataaccc
      661 tcgcctttat tttctttgat agctttatca tcaaatattt tgttattttt cttattcatc
      721 acacccagat aatatttatc atctttgata aataaaattg ccgtattgtc aggctcttta
      781 tttttatccc aaccattagc caaagtcgag ttctcaaaat tgagcttaaa tttctcatca
      841 ctatatggct tttgagttat atagtttcta attttgttat aaagaggcac tatattcgct
      901 agctcaaagt agcactcctc aaatactaga taaaaatgct catccttgtc taaaatattt
      961 gccttatctt ctgactgact aatatgaaat atttttagtt tatgtaagag attattagtt
     1021 tgatctaaaa gatccttgat agctttaaca tcatcttccg cactagcttg aagtaggtct
     1081 tttttacctt gattttgata tttgatagat atctgtgcca aattgtcttt gttttgagct
     1141 atttcatcaa atatcatcgg aatagccgca aagtttgcaa gtatttcttc aaacctacac
     1201 tgtttatcta tatctctatg cttattaaat tcttctaagg caagctttat agtttctaga
     1261 gataagtatt ttgctttttc agtttttttg gctattaatt cttgctcttt cttactaggg
     1321 ttatcaagat ttttaggtgc tatttgttga gttatatatt ctagtaccgc tgtaccaata
     1381 acactataat catcaaaaac ttgttgtgat agatcagtaa gagatttatc atttttaaaa
     1441 taaattttac tcaaatcaag tttttgagct tttaaatcat caaataataa agatagtgtt
     1501 tctttaatag atttttcttc tactgtttta aaagctgcta tttgctcata aaaactttgc
     1561 atcgttgtaa ctacatcact atcatcttct aacttatcaa ttacaaaaga tttagattct
     1621 gtatcactta aaatttgctt aaataaaaca ctcattttat attttttgag tgttttatca
     1681 tttatttgct gtgagtatag atttatatat tcatttatac cttttctctt tgtattttca
     1741 ccatttacaa atttaccacc aataatagta ttaaatttag taataccact ttgatttaga
     1801 taattattaa agtttgctat ctcaaaaact tcatcaagtg aaaaaactct ttgattaact
     1861 tcagatgttt tgtagtcaat atcaaaggtt agctcttctg ccaaatcttt tttaatttgt
     1921 tcatagttta tagcttctgg agctttgtct tttaaactct catacttagc tttattttct
     1981 agaaatttag gcaaattatc atctactatc ctataaataa tagatgtagg aatatcattg
     2041 ctactataaa cattttttct attttcatga aaacccttaa aataagttgt ccaaccttta
     2101 aaagatttga ttatttctaa cgcctcatct atatctgtga tatcactatt ggctttaaat
     2161 agttctatac cattatcctt agattgcttt agccatagaa ttaaatctga ctcttgccct
     2221 tttttagcat cgataaggtt ttgattaaac aaattcttaa atttctctga gtcctttata
     2281 tattcagata tttgtttctt tatcgtatct tttgcacttt taaaatcttt ttgtagatta
     2341 tcatcatcac tctttttaag tttaaaataa acatcagaat agttttgtaa taaatcttcg
     2401 ctaatacaaa ccgaacttaa tatctcctct ataaaaaact gatgatattt atcaattatt
     2461 tgtttagcct ttttgtagtc tttagctctt ttctcatcat ctaaaatcaa acctcttgct
     2521 tttatgtttt caagtgtttt accctgtggg attaactcaa atcttagagt tttacttaaa
     2581 ctatatttat taacaaattc ttgataaatt gacatagatc ctttctcctc ttttttattc
     2641 gactataaca aaccattttc ttgcgtaaac ctgtacgatc ctacaggtag atctttaatt
     2701 tttaaaaaat ggccaatcaa ttgctcctgt taaaattgct ttagaaatac tttggcagcg
     2761 gtttgttgta tcgagtttca tttgcgcatt ggttaaatgg aaagtgacag tacgctcact
     2821 gcagcctaat atttttgaaa tatcccaaga gctttttcct tcgcatgccc acgctaaaca
     2881 ttctttttct cttttggtta aatcgttgtt tgatttatta tttgctatat ttatttttcg
     2941 ataattatca actagagaag gaacaattaa tggtatgttc atacacgcat gtaaaaataa
     3001 actatctata tagttgtcaa tttctgaatg tgcaaaacta agcattccga agccattgtt
     3061 agccgtatga atagggaaac taaacccagt gataagacct gatgttttcg cttctttaat
     3121 tacatttgga gattttttat ttacagcatt gttttcaaat atattccaat taattggtga
     3181 atgattggat ttagaataat ctactatagg atcatatttt attaaattag cgcgccataa
     3241 tattgcctcc attttttagg gtaattatct agaattgaaa tatcagattt aaccatagaa
     3301 tgaggataaa tgatcgcgag taaataatat tcacaatgta ccattttagt catatcagat
     3361 aagcattgat taatatcatt attgctgcca caagctttaa ttttattaat tattctgtat
     3421 gtgtcgtcgg catttatgtt tttcataccc atctctttat ccttacctat tgtttgtcgc
     3481 aagttttgcg tgttatatat cattaaaacg gtaatggatt gacatttgat tctaataaat
     3541 tggatttttg tcacactatt gtatcgctgg gaatacaatt acttaacata agcacctgta
     3601 ggatcgtaca ggtttacgca agaaaatggt ttgttatagt cgaataaacg caagggaggt
     3661 tggtatgact ataatgataa aaaaatcgga ttttttggca attccatcgg aggagtataa
     3721 aggtattcta agtcttcgtt atcaagtgtt taagcaaaga cttgagtggg acttagttgt
     3781 agaaaataac cttgaatcag atgagtatga taactcaaat gcagaatata tttatgcttg
     3841 tgatgatact gaaaatgtaa gtggatgctg gcgtttatta cctacaacag gtgattatat
     3901 gctgaaaagt gtttttcctg aattgcttgg tcaacagagt gctcccaaag atcctaatat
     3961 agtcgaatta agtcgttttg ctgtaggtaa aaatagctca aagataaata actctgctag
     4021 tgaaattaca atgaaactat ttgaagctat atataaacac gctgttagtc aaggtattac
     4081 agaatatgta acagtaacat caacagcaat agagcgattt ttaaagcgta ttaaagttcc
     4141 ttgtcatcgt attggagaca aagaaattca tgtattaggt gatactaaat cggttgtatt
     4201 gtctatgcct attaatgaac agtttaaaaa agcagtctta aattaatcta gataagttca
     4261 ggatgaattc tctagtccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     4321 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     4381 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     4441 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     4501 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     4561 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     4621 cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     4681 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     4741 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     4801 gccgcattgc atcagccatg atggatactt tctcgacagg agcaaggtga gatgacagga
     4861 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     4921 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     4981 cctgtagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     5041 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     5101 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     5161 tcatgcgaaa cgatcctcat cctgtctctt gatcaaatct tgatcccctg cgccatcaga
     5221 tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg
     5281 gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc
     5341 gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc
     5401 agatagccca gtagctgaca ttcatcccag gtggcacttt tcggggaaat gtgcgcgccc
     5461 gcgttcctac tggcgctggg cctgtttctg gcgctggact tcccgctgtt ccgtcagcag
     5521 cttttcgccc acggccttga tgatcgcggc ggccttggcc tgcatatccc gattcaacgg
     5581 ccccagggcg tccagaacgg gcttcaggcg ctcccgaagg tctcgggccg tctcttgggc
     5641 ttgatcggcc ttcttgcgca tctcacgcgc tcctgcggcg gcctgtaggg caggctcata
     5701 cccctgccga accgcttttg tcagccggtc ggccacggct tccggcgtct caacgcgctt
     5761 tgagattccc agcttttcgg ccaatccctg cggtgcatag gcgcgtggct cgaccgcttg
     5821 cgggctgatg gtgacgtggc ccactggtgg ccgctccagg gcctcgtaga acgcctgaat
     5881 gcgcgtgtga cgtgccttgc tgccctcgat gccccgttgc agccctagat cggccacagc
     5941 agccgcaaac gtggtctggt cgcgggtcat ctgcgctttg ttgccgatga actccttggc
     6001 cgacagcctg ccgtcctgcg tcagcggcac cacgaacgcg gtcatgtgcg ggctggtttc
     6061 gtcacggtgg atgctggccg tcacgatgcg atccgccccg tacttgtccg ccagccactt
     6121 gtgcgccttc tcgaagaacg ccgcctgctg ttcttggctg gccgacttcc accattccgg
     6181 gctggccgtc atgacgtact cgaccgccaa cacagcgtcc ttgcgccgct tctctggcag
     6241 caactcgcgc agtcggccca tcgcttcatc ggtgctgctg gccgcccagt gctcgttctc
     6301 tggcgtcctg ctggcgtcag cgttgggcgt ctcgcgctcg cggtaggcgt gcttgagact
     6361 ggccgccacg ttgcccattt tcgccagctt cttgcatcgc atgatcgcgt atgccgccat
     6421 gcctgcccct cccttttggt gtccaaccgg ctcgacgggg gcagcgcaag gcggtgcctc
     6481 cggcgggcca ctcaatgctt gagtatactc actagacttt gcttcgcaaa gtcgtgaccg
     6541 cctacggcgg ctgcggcgcc ctacgggctt gctctccggg cttcgccctg cgcggtcgct
     6601 gcgctccctt gccagcccgt ggatatgtgg acgatggccg cgagcggcca ccggctggct
     6661 cgcttcgctc ggcccgtgga caaccctgct ggacaagctg atggacaggc tgcgcctgcc
     6721 cacgagcttg accacaggga ttgcccaccg gctacccagc cttcgaccac atacccaccg
     6781 gctccaactg cgcggcctgc ggccttgccc catcaatttt tttaattttc tctggggaaa
     6841 agcctccggc ctgcggcctg cgcgcttcgc ttgccggttg gacaccaagt ggaaggcggg
     6901 tcaaggctcg cgcagcgacc gcgcagcggc ttggccttga cgcgcctgga acgacccaag
     6961 cctatgcgag tgggggcagt cgaagggcga agcccgcccg cctgcccccc gagcctcacg
     7021 gcggcgagtg cgggggttcc aagggggcag cgccaccttg ggcaaggccg aaggccgcgc
     7081 agtcgatcaa caagccccgg aggggccact ttttgccgga gggggagccg cgccgaaggc
     7141 gtgggggaac cccgcagggg tgcccttctt tgggcaccaa agaactagat atagggcgaa
     7201 atgcgaaaga cttaaaaatc aacaacttaa aaaagggggg tacgcaacag ctcattgcgg
     7261 caccccccgc aatagctcat tgcgtaggtt aaagaaaatc tgtaattgac tgccactttt
     7321 acgcaacgca taattgttgt cgcgctgcca aaaagttgca gctgattgtg catggtgccg
     7381 caaccgtgcg gcacccctac cgcatggaga taagcatggc cacgcagtcc agagaaatcg
     7441 gcattcaagc caagaacaag cccggtcact gggtgcaaac ggaacgcaaa gcgcatgagg
     7501 cgtgggccgg gcttattgcg aggaaaccca cggcggcaat gctgctgcat cacctcgtgg
     7561 cgcagatggg ccaccagaac gccgtggtgg tcagccagaa gacactttcc aagctcatcg
     7621 gacgttcttt gcggacggtc caatacgcag tcaaggactt ggtggccgag cgctggatct
     7681 ccgtcgtgaa gctcaacggc cccggcaccg tgtcggccta cgtggtcaat gaccgcgtgg
     7741 cgtggggcca gccccgcgac cagttgcgcc tgtcggtgtt cagtgccgcc gtggtggttg
     7801 atcacgacga ccaggacgaa tcgctgttgg ggcatggcga cctgcgccgc atcccgaccc
     7861 tgtatccggg cgagtagcaa ctaccgaccg gccccggcga ggagccgccc agccagcccg
     7921 gcattccggg catggaacca gacctgccag ccttgaccga aacggaggaa tgggaacggc
     7981 gcgggcagca gcgcctgccg atgcccgatg agccgtgttt tctggacgat ggcgagccgt
     8041 tggagccgcc gacacgggtc acgctgccgc gccggtagca cttgggttgc gcagcaaccc
     8101 gtaagtgcgc tgttccagac tatactagac tgcaggtcct gaagttaaca aaaaaaatga
     8161 ccttcataaa tcgctaatct acaacagtag aaattattta aagttcttag accggctact
     8221 aatacaaaag gctctagaca tctacaacag tagaaattat ttaaagttct tagactgaat
     8281 tatctgaagg cacaggaata gtagcatcta caacagtaga aattatttaa agttcttaga
     8341 cgctcctcgc ccttgctcac catgctcgag ccctgatcta caacagtaga aattatttaa
     8401 agttcttaga cccgtttttg cctaaatcag cgctagcatt atacctagga ctgagctagc
     8461 tgtcaactcg agttagttat tcctattctg cacgaactca aaatactctt catttttgat
     8521 aaccaaattg agttttttgc cctcttgatt atttttgatc ctacctagta gcatcagacc
     8581 ttttagccca atatgataag caccattggc atcagcatct tgaggcatat tttttggcgc
     8641 ctgtcgcgaa tcaaagaaat tgccatttac atctgctact ggtgaaatta gataatctaa
     8701 ctcagtacct gtttttgagt tacgcatttg taagatagta tttaggacac tagttagctt
     8761 agcaaaaaac tttttgtcgc tctcaccgca aatagctgct ttgatacatt cgccatgccc
     8821 atattcgata gaataatctt ttagcaattt ctccaactct ttagttggat aaacttctcg
     8881 agtatcccaa ttatgatttt tatctgaatt tctaaagtta atcaatctac tcccaaagct
     8941 agctatagtc cacttgcctt tggcagcctt gtcaccaaag tttttataat caaaactaaa
     9001 ctcaaaatag cccttatcaa ggttataaca aatcttgtca aacttactaa agaactcttg
     9061 agatttgctg acactttcat acttaggata taactgattt acaaaaccag ttacaggaca
     9121 aatttttgaa gtaaaaccag ctggtacata gtagataata cctgtttgtt tacccatctt
     9181 tttaaaagtc tcaaaaggtg ctgttagctg ataagctcta agcactcccc cagttttatc
     9241 aaactcatta tctttgaaaa ctagatagtt tagtttctca attagcattt tttctaactt
     9301 ttgatagacc tgcttctcta ccttgaaacg ccctctttta aatccaaaat ttaaatcctc
     9361 aaaaaccaca atagcattat actctataac tagcttagct atttcatgaa ctacctgaga
     9421 tagatagccc tctttcatct ctttgatgtt atttatcttt ttccagtctt tcctagctga
     9481 atccctatct ttctctattg cagcaagctt atcatggtag tttgttttca ttctatcatt
     9541 accaatgatg ttgaaagtat cttgtttgat gatattgcct ttaccatcta ccaaagtata
     9601 gtaagctaaa tgtctttcac ctctagctat acttaatata tgaacatcat ttgctttttc
     9661 ttttagcaat aaattgattt catcattaaa cttattagct ccactagatt taaaattgat
     9721 tgtaatagga cagtgaaaga aaaacttatc ttcagtaaag
//
