LOCUS       sequence-469925-        9761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid pQdCas12a.sggfp(C) expresses the dCas12a endonuclease 
            and the sgRNA (design C) targeting the gfp gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9761)
  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 9761)
  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.sggfp(C)
FEATURES             Location/Qualifiers
     source          1..9761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      54..825
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             826..1488
                     /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(1842..1876)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             5792..5803
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(7541..8335)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7652..7671)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8262..8281
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 cgcggtcgct gcgctccctt gccagcccgt ggatatgtgg acgatggccg cgagcggcca
       61 ccggctggct cgcttcgctc ggcccgtgga caaccctgct ggacaagctg atggacaggc
      121 tgcgccagcc cacgagcttg accacaggga ttgcccaccg gctacccagc cttcgaccac
      181 atacccaccg gctccaactg cgcggcctgc ggccttgccc catcaatttt tttaattttc
      241 tctggggaaa agcctccggc ctgcggcctg cgcgcttcgc ttgccggttg gacaccaagt
      301 ggaaggcggg tcaaggctcg cgcagcgacc gcgcagcggc ttggccttga cgcgcctgga
      361 acgacccaag cctatgcgag tgggggcagt cgaagggcga agcccgcccg cctgcccccc
      421 gagcctcacg gcggcgagtg cgggggttcc aagggggcag cgccaccttg ggcaaggccg
      481 aaggccgcgc agtcgatcaa caagccccgg aggggccact ttttgccgga gggggagccg
      541 cgccgaaggc gtgggggaac cccgcagggg tgcccttctt tgggcaccaa agaactagat
      601 atagggcgaa atgcgaaaga cttaaaaatc aacaacttaa aaaagggggg tacgcaacag
      661 ctcattgcgg caccccccgc aatagctcat tgcgtaggtt aaagaaaatc tgtaattgac
      721 tgccactttt acgcaacgca taattgttgt cgcgctgcca aaaagttgca gctgattgcg
      781 catggtgccg caaccgtgcg gcacccctac cgcatggaga taagcatggc cacgcagtcc
      841 agagaaatcg gcattcaagc caagaacaag cccggtcact gggtgcaaac ggaacgcaaa
      901 gcgcatgagg cgtgggccgg gcttattgcg aggaaaccca cggcggcaat gctgctgcat
      961 cacctcgtgg cgcagatggg ccaccagaac gccgtggtgg tcagccagaa gacactttcc
     1021 aagctcatcg gacgttcttt gcggacggtc caatacgcag tcaaggactt ggtggccgag
     1081 cgctggatct ccgtcgtgaa gctcaacggc cccggcaccg tgtcggccta cgtggtcaat
     1141 gaccgcgtgg cgtggggcca gccccgcgac cagttgcgcc tgtcggtgtt cagtgccgcc
     1201 gtggtggttg atcacgacga ccaggacgaa tcgctgttgg ggcatggcga cctgcgccgc
     1261 atcccgaccc tgtatccggg cgagtagcaa ctaccgaccg gccccggcga ggagccgccc
     1321 agccagcccg gcattccggg catggaacca gacctgccag ccttgaccga aacggaggaa
     1381 tgggaacggc gcgggcagca gcgcctgccg atgcccgatg agccgtgttt tctggacgat
     1441 ggcgagccgt tggagccgcc gacacgggtc acgctgccgc gccggtagca cttgggttgc
     1501 gcagcaaccc gtaagtgcgc tgttccagac tatactagac tgcaggtcct gaagttaaca
     1561 aaaaaaatga ccttcataaa tcgctaatct acaacagtag aaattattta aagttcttag
     1621 accggctact aatacaaaag gctctagaca tctacaacag tagaaattat ttaaagttct
     1681 tagactgaat tatctgaagg cacaggaata gtagcatcta caacagtaga aattatttaa
     1741 agttcttaga cagaacctgt acttccaggg ctcgagcatg gtgagatcta caacagtaga
     1801 aattatttaa agttcttaga cccgtttttg cctaaatcag cgctagcatt atacctagga
     1861 ctgagctagc tgtcaactcg agttagttat tcctattctg cacgaactca aaatactctt
     1921 catttttgat aaccaaattg agttttttgc cctcttgatt atttttgatc ctacctagta
     1981 gcatcagacc ttttagccca atatgataag caccattggc atcagcatct tgaggcatat
     2041 tttttggcgc ctgtcgcgaa tcaaagaaat tgccatttac atctgctact ggtgaaatta
     2101 gataatctaa ctcagtacct gtttttgagt tacgcatttg taagatagta tttaggacac
     2161 tagttagctt agcaaaaaac tttttgtcgc tctcaccgca aatagctgct ttgatacatt
     2221 cgccatgccc atattcgata gaataatctt ttagcaattt ctccaactct ttagttggat
     2281 aaacttctcg agtatcccaa ttatgatttt tatctgaatt tctaaagtta atcaatctac
     2341 tcccaaagct agctatagtc cacttgcctt tggcagcctt gtcaccaaag tttttataat
     2401 caaaactaaa ctcaaaatag cccttatcaa ggttataaca aatcttgtca aacttactaa
     2461 agaactcttg agatttgctg acactttcat acttaggata taactgattt acaaaaccag
     2521 ttacaggaca aatttttgaa gtaaaaccag ctggtacata gtagataata cctgtttgtt
     2581 tacccatctt tttaaaagtc tcaaaaggtg ctgttagctg ataagctcta agcactcccc
     2641 cagttttatc aaactcatta tctttgaaaa ctagatagtt tagtttctca attagcattt
     2701 tttctaactt ttgatagacc tgcttctcta ccttgaaacg ccctctttta aatccaaaat
     2761 ttaaatcctc aaaaaccaca atagcattat actctataac tagcttagct atttcatgaa
     2821 ctacctgaga tagatagccc tctttcatct ctttgatgtt atttatcttt ttccagtctt
     2881 tcctagctga atccctatct ttctctattg cagcaagctt atcatggtag tttgttttca
     2941 ttctatcatt accaatgatg ttgaaagtat cttgtttgat gatattgcct ttaccatcta
     3001 ccaaagtata gtaagctaaa tgtctttcac ctctagctat acttaatata tgaacatcat
     3061 ttgctttttc ttttagcaat aaattgattt catcattaaa cttattagct ccactagatt
     3121 taaaattgat tgtaatagga cagtgaaaga aaaacttatc ttcagtaaag cgtttatctt
     3181 tgattaaatc atattcaaaa acactctctt ttttaggatt atctttgttt ttattagcta
     3241 ttgcctcttt agctgggtga gtgatttttt taggtattga ttgtttacga taaaaaagct
     3301 ctgcctcacc atttagctta taaaccacat cttgaagatt tctctcatca aacagcgctt
     3361 tccaatataa agtatgtaga tttggtcgcc ctttgctata agctgaaaaa tctttattat
     3421 agatttggaa taggtacaat ttaccctgat taactacgct atcaatatag ctctctgata
     3481 tattttcaaa agttagtttg tagccttgat tttcaacttc tctataaaat tcatctatag
     3541 aattatatct ttgagtatca gaaaatctaa atccaaaatc tttccactcc ggatgcttac
     3601 ttatagactg tttataaaaa tctataaatt ttcggcaatc ttcaatatta aactcaaatt
     3661 tttcatatcc tttttgagga ctaccatttt ttgtatgtgt ggaatgattt cttattctaa
     3721 gtatatcttc actaggatta taaaatttta tagatttagc agaaaagaaa accttaggta
     3781 acattttatt tgcgccaggt aaaagtttat aaacaatttt tttataaccc tcgcctttat
     3841 tttctttgat agctttatca tcaaatattt tgttattttt cttattcatc acacccagat
     3901 aatatttatc atctttgata aataaaattg ccgtattgtc aggctcttta tttttatccc
     3961 aaccattagc caaagtcgag ttctcaaaat tgagcttaaa tttctcatca ctatatggct
     4021 tttgagttat atagtttcta attttgttat aaagaggcac tatattcgct agctcaaagt
     4081 agcactcctc aaatactaga taaaaatgct catccttgtc taaaatattt gccttatctt
     4141 ctgactgact aatatgaaat atttttagtt tatgtaagag attattagtt tgatctaaaa
     4201 gatccttgat agctttaaca tcatcttccg cactagcttg aagtaggtct tttttacctt
     4261 gattttgata tttgatagat atctgtgcca aattgtcttt gttttgagct atttcatcaa
     4321 atatcatcgg aatagccgca aagtttgcaa gtatttcttc aaacctacac tgtttatcta
     4381 tatctctatg cttattaaat tcttctaagg caagctttat agtttctaga gataagtatt
     4441 ttgctttttc agtttttttg gctattaatt cttgctcttt cttactaggg ttatcaagat
     4501 ttttaggtgc tatttgttga gttatatatt ctagtaccgc tgtaccaata acactataat
     4561 catcaaaaac ttgttgtgat agatcagtaa gagatttatc atttttaaaa taaattttac
     4621 tcaaatcaag tttttgagct tttaaatcat caaataataa agatagtgtt tctttaatag
     4681 atttttcttc tactgtttta aaagctgcta tttgctcata aaaactttgc atcgttgtaa
     4741 ctacatcact atcatcttct aacttatcaa ttacaaaaga tttagattct gtatcactta
     4801 aaatttgctt aaataaaaca ctcattttat attttttgag tgttttatca tttatttgct
     4861 gtgagtatag atttatatat tcatttatac cttttctctt tgtattttca ccatttacaa
     4921 atttaccacc aataatagta ttaaatttag taataccact ttgatttaga taattattaa
     4981 agtttgctat ctcaaaaact tcatcaagtg aaaaaactct ttgattaact tcagatgttt
     5041 tgtagtcaat atcaaaggtt agctcttctg ccaaatcttt tttaatttgt tcatagttta
     5101 tagcttctgg agctttgtct tttaaactct catacttagc tttattttct agaaatttag
     5161 gcaaattatc atctactatc ctataaataa tagatgtagg aatatcattg ctactataaa
     5221 cattttttct attttcatga aaacccttaa aataagttgt ccaaccttta aaagatttga
     5281 ttatttctaa cgcctcatct atatctgtga tatcactatt ggctttaaat agttctatac
     5341 cattatcctt agattgcttt agccatagaa ttaaatctga ctcttgccct tttttagcat
     5401 cgataaggtt ttgattaaac aaattcttaa atttctctga gtcctttata tattcagata
     5461 tttgtttctt tatcgtatct tttgcacttt taaaatcttt ttgtagatta tcatcatcac
     5521 tctttttaag tttaaaataa acatcagaat agttttgtaa taaatcttcg ctaatacaaa
     5581 ccgaacttaa tatctcctct ataaaaaact gatgatattt atcaattatt tgtttagcct
     5641 ttttgtagtc tttagctctt ttctcatcat ctaaaatcaa acctcttgct tttatgtttt
     5701 caagtgtttt accctgtggg attaactcaa atcttagagt tttacttaaa ctatatttat
     5761 taacaaattc ttgataaatt gacatagatc ctttctcctc ttttttattc gactataaca
     5821 aaccattttc ttgcgtaaac ctgtacgatc ctacaggtag atctttaatt tttaaagtat
     5881 gggcaatcaa ttgctcctgt taaaattgct ttagaaatac tttggcagcg gtttgttgta
     5941 ttgagtttca tttgcgcatt ggttaaatgg aaagtgacag tacgctcact gcagcctaat
     6001 atttttgaaa tatcccaaga gctttttcct tcgcatgccc acgctaaaca ttctttttct
     6061 cttttggtta aatcgttgtt tgatttatta tttgctatat ttatttttcg ataattatca
     6121 actagagaag gaacaattaa tggtatgttc atacacgcat gtaaaaataa actatctata
     6181 tagttgtctt tttctgaatg tgcaaaacta agcattccga agccattgtt agccgtatga
     6241 atagggaaac taaacccagt gataagacct gatgttttcg cttctttaat tacatttgga
     6301 gattttttat ttacagcatt gttttcaaat atattccaat taattggtga atgattggag
     6361 ttagaataat ctactatagg atcatatttt attaaattag cgtcatcata atattgcctc
     6421 cattttttag ggtaattatc tagaattgaa atatcagatt taaccataga atgaggataa
     6481 atgatcgcga gtaaataata ttcacaatgt accattttag tcatatcaga taagcattga
     6541 ttaatatcat tattgcttct acaagcttta attttattaa ttattctgta tgtgtcgtcg
     6601 gcatttatgt ttttcatacc catctcttta tccttaccta ttgtttgtcg caagttttgc
     6661 gtgttatata tcattaaaac ggtaatggat tgacatttga ttctaataaa ttggattttt
     6721 gtcacactat tgtatcgctg ggaatacaat tacttaacat aagcacctgt aggatcgtac
     6781 aggtttacgc aagaaaatgg tttgttatag tcgaataaac gcaagggagg ttggtatgac
     6841 tataatgata aaaaaatcgg attttttggc aattccatcg gaggagtata aaggtattct
     6901 aagtcttcgt tatcaagtgt ttaagcaaag acttgagtgg gacttagttg tagaaaataa
     6961 ccttgaatca gatgagtatg ataactcaaa tgcagaatat atttatgctt gtgatgatac
     7021 tgaaaatgta agtggatgct ggcgtttatt acctacaaca ggtgattata tgctgaaaag
     7081 tgtttttcct gaattgcttg gtcaacagag tgctcccaaa gatcctaata tagtcgaatt
     7141 aagtcgtttt gctgtaggta aaaatagctc aaagataaat aactctgcta gtgaaattac
     7201 aatgaaacta tttgaagcta tatataaaca cgctgttagt caaggtatta cagaatatgt
     7261 aacagtaaca tcaacagcaa tagagcgatt tttaaagcgt attaaagttc cttgtcatcg
     7321 tattggagac aaagaaattc atgtattagg tgatactaaa tcggttgtat tgtctatgcc
     7381 tattaatgaa cagtttaaaa aagcagtctt aaattaatct agataagttc aggatgaatt
     7441 ctctagtcca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
     7501 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     7561 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     7621 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     7681 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     7741 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     7801 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     7861 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     7921 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     7981 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     8041 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     8101 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgtagtt
     8161 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     8221 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     8281 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     8341 acgatcctca tcctgtctct tgatcaaatc ttgatcccct gcgccatcag atccttggcg
     8401 gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag
     8461 ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
     8521 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
     8581 agtagctgac attcatccca ggtggcactt ttcggggaaa tgtgcgcgcc cgcgttcctg
     8641 ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt tccgtcagca gcttttcgcc
     8701 cacggccttg atgatcgcgg cggccttggc ctgcatatcc cgattcaacg gccccagggc
     8761 gtccagaacg ggcttcaggc gctcccgaag gtctcgggcc gtctcttggg cttgatcggc
     8821 cttcttgcgc atctcacgcg ctcctgcggc ggcctgtagg gcaggctcat acccctgccg
     8881 aaccgctttt gtcagccggt cggccacggc ttccggcgtc tcaacgcgct ttgagattcc
     8941 cagcttttcg gccaatccct gcggtgcata ggcgcgtggc tcgaccgctt gcgggctgat
     9001 ggtgacgtgg cccactggtg gccgctccag ggcctcgtag aacgcctgaa tgcgcgtgtg
     9061 acgtgccttg ctgccctcga tgccccgttg cagccctaga tcggccacag cagccgcaaa
     9121 cgtggtctgg tcgcgggtca tctgcgcttt gttgccgatg aactccttgg ccgacagcct
     9181 gccgtcctgc gtcagcggca ccacgaacgc ggtcatgtgc gggctggttt cgtcacggtg
     9241 gatgctggcc gtcacgatgc gatccgcccc gtacttgtcc gccagccact tgtgcgcctt
     9301 ctcgaagaac gccgcctgct gttcttggct ggccgacttc caccattccg ggctggccgt
     9361 catgacgtac tcgaccgcca acacagcgtc cttgcgccgc ttctctggca gcaactcgcg
     9421 cagtcggccc atcgcttcat cggtgctgct ggccgcccag tgctcgttct ctggcgtcct
     9481 gctggcgtca gcgttgggcg tctcgcgctc gcggtaggcg tgcttgagac tggccgccac
     9541 gttgcccatt ttcgccagct tcttgcatcg catgatcgcg tatgccgcca tgcctgcccc
     9601 tcccttttgg tgtccaaccg gctcgacggg ggcagcgcaa ggcggtgcct ccggcgggcc
     9661 actcaatgct tgagtatact cactagactt tgcttcgcaa agtcgtgacc gcctacggcg
     9721 gctgcggcgc cctacgggct tgctctccgg gcttcgccct g
//
