LOCUS       sequence-471082-        9759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid pQdCas12a.luxR(mut)-sggfp(C) expresses the dCas12a 
            endonuclease and the sgRNA (design C) 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 9759)
  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 9759)
  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(C)
FEATURES             Location/Qualifiers
     source          1..9759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1189..1960
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1961..2623
                     /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(2976..3010)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             6926..6937
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(8674..9468)
                     /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(8785..8804)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     9395..9414
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 cgcccgcgtt cctactggcg ctgggcctgt ttctggcgct ggacttcccg ctgttccgtc
       61 agcagctttt cgcccacggc cttgatgatc gcggcggcct tggcctgcat atcccgattc
      121 aacggcccca gggcgtccag aacgggcttc aggcgctccc gaaggtctcg ggccgtctct
      181 tgggcttgat cggccttctt gcgcatctca cgcgctcctg cggcggcctg tagggcaggc
      241 tcatacccct gccgaaccgc ttttgtcagc cggtcggcca cggcttccgg cgtctcaacg
      301 cgctttgaga ttcccagctt ttcggccaat ccctgcggtg cataggcgcg tggctcgacc
      361 gcttgcgggc tgatggtgac gtggcccact ggtggccgct ccagggcctc gtagaacgcc
      421 tgaatgcgcg tgtgacgtgc cttgctgccc tcgatgcccc gttgcagccc tagatcggcc
      481 acagcagccg caaacgtggt ctggtcgcgg gtcatctgcg ctttgttgcc gatgaactcc
      541 ttggccgaca gcctgccgtc ctgcgtcagc ggcaccacga acgcggtcat gtgcgggctg
      601 gtttcgtcac ggtggatgct ggccgtcacg atgcgatccg ccccgtactt gtccgccagc
      661 cacttgtgcg ccttctcgaa gaacgccgcc tgctgttctt ggctggccga cttccaccat
      721 tccgggctgg ccgtcatgac gtactcgacc gccaacacag cgtccttgcg ccgcttctct
      781 ggcagcaact cgcgcagtcg gcccatcgct tcatcggtgc tgctggccgc ccagtgctcg
      841 ttctctggcg tcctgctggc gtcagcgttg ggcgtctcgc gctcgcggta ggcgtgcttg
      901 agactggccg ccacgttgcc cattttcgcc agcttcttgc atcgcatgat cgcgtatgcc
      961 gccatgcctg cccctccctt ttggtgtcca accggctcga cgggggcagc gcaaggcggt
     1021 gcctccggcg ggccactcaa tgcttgagta tactcactag actttgcttc gcaaagtcgt
     1081 gaccgcctac ggcggctgcg gcgccctacg ggcttgctct ccgggcttcg ccctgcgcgg
     1141 tcgctgcgct cccttgccag cccgtggata tgtggacgat ggccgcgagc ggccaccggc
     1201 tggctcgctt cgctcggccc gtggacaacc ctgctggaca agctgatgga caggctgcgc
     1261 ctgcccacga gcttgaccac agggattgcc caccggctac ccagccttcg accacatacc
     1321 caccggctcc aactgcgcgg cctgcggcct tgccccatca atttttttaa ttttctctgg
     1381 ggaaaagcct ccggcctgcg gcctgcgcgc ttcgcttgcc ggttggacac caagtggaag
     1441 gcgggtcaag gctcgcgcag cgaccgcgca gcggcttggc cttgacgcgc ctggaacgac
     1501 ccaagcctat gcgagtgggg gcagtcgaag ggcgaagccc gcccgcctgc cccccgagcc
     1561 tcacggcggc gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa ggccgaaggc
     1621 cgcgcagtcg atcaacaagc cccggagggg ccactttttg ccggaggggg agccgcgccg
     1681 aaggcgtggg ggaaccccgc aggggtgccc ttctttgggc accaaagaac tagatatagg
     1741 gcgaaatgcg aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc aacagctcat
     1801 tgcggcaccc cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa ttgactgcca
     1861 cttttacgca acgcataatt gttgtcgcgc tgccaaaaag ttgcagctga ttgtgcatgg
     1921 tgccgcaacc gtgcggcacc cctaccgcat ggagataagc atggccacgc agtccagaga
     1981 aatcggcatt caagccaaga acaagcccgg tcactgggtg caaacggaac gcaaagcgca
     2041 tgaggcgtgg gccgggctta ttgcgaggaa acccacggcg gcaatgctgc tgcatcacct
     2101 cgtggcgcag atgggccacc agaacgccgt ggtggtcagc cagaagacac tttccaagct
     2161 catcggacgt tctttgcgga cggtccaata cgcagtcaag gacttggtgg ccgagcgctg
     2221 gatctccgtc gtgaagctca acggccccgg caccgtgtcg gcctacgtgg tcaatgaccg
     2281 cgtggcgtgg ggccagcccc gcgaccagtt gcgcctgtcg gtgttcagtg ccgccgtggt
     2341 ggttgatcac gacgaccagg acgaatcgct gttggggcat ggcgacctgc gccgcatccc
     2401 gaccctgtat ccgggcgagt agcaactacc gaccggcccc ggcgaggagc cgcccagcca
     2461 gcccggcatt ccgggcatgg aaccagacct gccagccttg accgaaacgg aggaatggga
     2521 acggcgcggg cagcagcgcc tgccgatgcc cgatgagccg tgttttctgg acgatggcga
     2581 gccgttggag ccgccgacac gggtcacgct gccgcgccgg tagcacttgg gttgcgcagc
     2641 aacccgtaag tgcgctgttc cagactatac tagactgcag gtcctgaagt taacaaaaaa
     2701 aatgaccttc ataaatcgct aatctacaac agtagaaatt atttaaagtt cttagaccgg
     2761 ctactaatac aaaaggctct agacatctac aacagtagaa attatttaaa gttcttagac
     2821 tgaattatct gaaggcacag gaatagtagc atctacaaca gtagaaatta tttaaagttc
     2881 tagacagaac ctgtacttcc agggctcgag catggtgaga tctacaacag tagaaattat
     2941 ttaaagttct tagacccgtt tttgcctaaa tcagcgctag cattatacct aggactgagc
     3001 tagctgtcaa ctcgagttag ttattcctat tctgcacgaa ctcaaaatac tcttcatttt
     3061 tgataaccaa attgagtttt ttgccctctt gattattttt gatcctacct agtagcatca
     3121 gaccttttag cccaatatga taagcaccat tggcatcagc atcttgaggc atattttttg
     3181 gcgcctgtcg cgaatcaaag aaattgccat ttacatctgc tactggtgaa attagataat
     3241 ctaactcagt acctgttttt gagttacgca tttgtaagat agtatttagg acactagtta
     3301 gcttagcaaa aaactttttg tcgctctcac cgcaaatagc tgctttgata cattcgccat
     3361 gcccatattc gatagaataa tcttttagca atttctccaa ctctttagtt ggataaactt
     3421 ctcgagtatc ccaattatga tttttatctg aatttctaaa gttaatcaat ctactcccaa
     3481 agctagctat agtccacttg cctttggcag ccttgtcacc aaagttttta taatcaaaac
     3541 taaactcaaa atagccctta tcaaggttat aacaaatctt gtcaaactta ctaaagaact
     3601 cttgagattt gctgacactt tcatacttag gatataactg atttacaaaa ccagttacag
     3661 gacaaatttt tgaagtaaaa ccagctggta catagtagat aatacctgtt tgtttaccca
     3721 tctttttaaa agtctcaaaa ggtgctgtta gctgataagc tctaagcact cccccagttt
     3781 tatcaaactc attatctttg aaaactagat agtttagttt ctcaattagc attttttcta
     3841 acttttgata gacctgcttc tctaccttga aacgccctct tttaaatcca aaatttaaat
     3901 cctcaaaaac cacaatagca ttatactcta taactagctt agctatttca tgaactacct
     3961 gagatagata gccctctttc atctctttga tgttatttat ctttttccag tctttcctag
     4021 ctgaatccct atctttctct attgcagcaa gcttatcatg gtagtttgtt ttcattctat
     4081 cattaccaat gatgttgaaa gtatcttgtt tgatgatatt gcctttacca tctaccaaag
     4141 tatagtaagc taaatgtctt tcacctctag ctatacttaa tatatgaaca tcatttgctt
     4201 tttcttttag caataaattg atttcatcat taaacttatt agctccacta gatttaaaat
     4261 tgattgtaat aggacagtga aagaaaaact tatcttcagt aaagcgttta tctttgatta
     4321 aatcatattc aaaaacactc tcttttttag gattatcttt gtttttatta gctattgcct
     4381 ctttagctgg gtgagtgatt tttttaggta ttgattgttt acgataaaaa agctctgcct
     4441 caccatttag cttataaacc acatcttgaa gatttctctc atcaaacagc gctttccaat
     4501 ataaagtatg tagatttggt cgccctttgc tataagctga aaaatcttta ttatagattt
     4561 ggaataggta caatttaccc tgattaacta cgctatcaat atagctctct gatatatttt
     4621 caaaagttag tttgtagcct tgattttcaa cttctctata aaattcatct atagaattat
     4681 atctttgagt atcagaaaat ctaaatccaa aatctttcca ctccggatgc ttacttatag
     4741 actgtttata aaaatctata aattttcggc aatcttcaat attaaactca aatttttcat
     4801 atcctttttg aggactacca ttttttgtat gtgtggaatg atttcttatt ctaagtatat
     4861 cttcactagg attataaaat tttatagatt tagcagaaaa gaaaacctta ggtaacattt
     4921 tatttgcgcc aggtaaaagt ttataaacaa tttttttata accctcgcct ttattttctt
     4981 tgatagcttt atcatcaaat attttgttat ttttcttatt catcacaccc agataatatt
     5041 tatcatcttt gataaataaa attgccgtat tgtcaggctc tttattttta tcccaaccat
     5101 tagccaaagt cgagttctca aaattgagct taaatttctc atcactatat ggcttttgag
     5161 ttatatagtt tctaattttg ttataaagag gcactatatt cgctagctca aagtagcact
     5221 cctcaaatac tagataaaaa tgctcatcct tgtctaaaat atttgcctta tcttctgact
     5281 gactaatatg aaatattttt agtttatgta agagattatt agtttgatct aaaagatcct
     5341 tgatagcttt aacatcatct tccgcactag cttgaagtag gtctttttta ccttgatttt
     5401 gatatttgat agatatctgt gccaaattgt ctttgttttg agctatttca tcaaatatca
     5461 tcggaatagc cgcaaagttt gcaagtattt cttcaaacct acactgttta tctatatctc
     5521 tatgcttatt aaattcttct aaggcaagct ttatagtttc tagagataag tattttgctt
     5581 tttcagtttt tttggctatt aattcttgct ctttcttact agggttatca agatttttag
     5641 gtgctatttg ttgagttata tattctagta ccgctgtacc aataacacta taatcatcaa
     5701 aaacttgttg tgatagatca gtaagagatt tatcattttt aaaataaatt ttactcaaat
     5761 caagtttttg agcttttaaa tcatcaaata ataaagatag tgtttcttta atagattttt
     5821 cttctactgt tttaaaagct gctatttgct cataaaaact ttgcatcgtt gtaactacat
     5881 cactatcatc ttctaactta tcaattacaa aagatttaga ttctgtatca cttaaaattt
     5941 gcttaaataa aacactcatt ttatattttt tgagtgtttt atcatttatt tgctgtgagt
     6001 atagatttat atattcattt ataccttttc tctttgtatt ttcaccattt acaaatttac
     6061 caccaataat agtattaaat ttagtaatac cactttgatt tagataatta ttaaagtttg
     6121 ctatctcaaa aacttcatca agtgaaaaaa ctctttgatt aacttcagat gttttgtagt
     6181 caatatcaaa ggttagctct tctgccaaat cttttttaat ttgttcatag tttatagctt
     6241 ctggagcttt gtcttttaaa ctctcatact tagctttatt ttctagaaat ttaggcaaat
     6301 tatcatctac tatcctataa ataatagatg taggaatatc attgctacta taaacatttt
     6361 ttctattttc atgaaaaccc ttaaaataag ttgtccaacc tttaaaagat ttgattattt
     6421 ctaacgcctc atctatatct gtgatatcac tattggcttt aaatagttct ataccattat
     6481 ccttagattg ctttagccat agaattaaat ctgactcttg ccctttttta gcatcgataa
     6541 ggttttgatt aaacaaattc ttaaatttct ctgagtcctt tatatattca gatatttgtt
     6601 tctttatcgt atcttttgca cttttaaaat ctttttgtag attatcatca tcactctttt
     6661 taagtttaaa ataaacatca gaatagtttt gtaataaatc ttcgctaata caaaccgaac
     6721 ttaatatctc ctctataaaa aactgatgat atttatcaat tatttgttta gcctttttgt
     6781 agtctttagc tcttttctca tcatctaaaa tcaaacctct tgcttttatg ttttcaagtg
     6841 ttttaccctg tgggattaac tcaaatctta gagttttact taaactatat ttattaacaa
     6901 attcttgata aattgacata gatcctttct cctctttttt attcgactat aacaaaccat
     6961 tttcttgcgt aaacctgtac gatcctacag gtagatcttt aatttttaaa aaatggccaa
     7021 tcaattgctc ctgttaaaat tgctttagaa atactttggc agcggtttgt tgtatcgagt
     7081 ttcatttgcg cattggttaa atggaaagtg acagtacgct cactgcagcc taatattttt
     7141 gaaatatccc aagagctttt tccttcgcat gcccacgcta aacattcttt ttctcttttg
     7201 gttaaatcgt tgtttgattt attatttgct atatttattt ttcgataatt atcaactaga
     7261 gaaggaacaa ttaatggtat gttcatacac gcatgtaaaa ataaactatc tatatagttg
     7321 tcaatttctg aatgtgcaaa actaagcatt ccgaagccat tgttagccgt atgaataggg
     7381 aaactaaacc cagtgataag acctgatgtt ttcgcttctt taattacatt tggagatttt
     7441 ttatttacag cattgttttc aaatatattc caattaattg gtgaatgatt ggatttagaa
     7501 taatctacta taggatcata ttttattaaa ttagcgcgcc ataatattgc ctccattttt
     7561 tagggtaatt atctagaatt gaaatatcag atttaaccat agaatgagga taaatgatcg
     7621 cgagtaaata atattcacaa tgtaccattt tagtcatatc agataagcat tgattaatat
     7681 cattattgct gccacaagct ttaattttat taattattct gtatgtgtcg tcggcattta
     7741 tgtttttcat acccatctct ttatccttac ctattgtttg tcgcaagttt tgcgtgttat
     7801 atatcattaa aacggtaatg gattgacatt tgattctaat aaattggatt tttgtcacac
     7861 tattgtatcg ctgggaatac aattacttaa cataagcacc tgtaggatcg tacaggttta
     7921 cgcaagaaaa tggtttgtta tagtcgaata aacgcaaggg aggttggtat gactataatg
     7981 ataaaaaaat cggatttttt ggcaattcca tcggaggagt ataaaggtat tctaagtctt
     8041 cgttatcaag tgtttaagca aagacttgag tgggacttag ttgtagaaaa taaccttgaa
     8101 tcagatgagt atgataactc aaatgcagaa tatatttatg cttgtgatga tactgaaaat
     8161 gtaagtggat gctggcgttt attacctaca acaggtgatt atatgctgaa aagtgttttt
     8221 cctgaattgc ttggtcaaca gagtgctccc aaagatccta atatagtcga attaagtcgt
     8281 tttgctgtag gtaaaaatag ctcaaagata aataactctg ctagtgaaat tacaatgaaa
     8341 ctatttgaag ctatatataa acacgctgtt agtcaaggta ttacagaata tgtaacagta
     8401 acatcaacag caatagagcg atttttaaag cgtattaaag ttccttgtca tcgtattgga
     8461 gacaaagaaa ttcatgtatt aggtgatact aaatcggttg tattgtctat gcctattaat
     8521 gaacagttta aaaaagcagt cttaaattaa tctagataag ttcaggatga attctctagt
     8581 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
     8641 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     8701 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     8761 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     8821 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     8881 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
     8941 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     9001 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     9061 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     9121 catgatggat actttctcga caggagcaag gtgagatgac aggagatcct gccccggcac
     9181 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     9241 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgta gttcattcag
     9301 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     9361 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     9421 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     9481 tcatcctgtc tcttgatcaa atcttgatcc cctgcgccat cagatccttg gcggcaagaa
     9541 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     9601 ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact
     9661 gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct
     9721 gacattcatc ccaggtggca cttttcgggg aaatgtgcg
//
