LOCUS       sequence-469933-        9761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid pQdCas12a.sggfp(A) expresses the dCas12a endonuclease 
            and the sgRNA (design A) 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(A)
FEATURES             Location/Qualifiers
     source          1..9761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             2296..2307
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(4045..4839)
                     /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(4156..4175)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4766..4785
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      6319..7090
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             7091..7753
                     /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(8107..8141)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 tttgtagcct tgattttcaa cttctctata aaattcatct atagaattat atctttgagt
       61 atcagaaaat ctaaatccaa aatctttcca ctccggatgc ttacttatag actgtttata
      121 aaaatctata aattttcggc aatcttcaat attaaactca aatttttcat atcctttttg
      181 aggactacca ttttttgtat gtgtggaatg atttcttatt ctaagtatat cttcactagg
      241 attataaaat tttatagatt tagcagaaaa gaaaacctta ggtaacattt tatttgcgcc
      301 aggtaaaagt ttataaacaa tttttttata accctcgcct ttattttctt tgatagcttt
      361 atcatcaaat attttgttat ttttcttatt catcacaccc agataatatt tatcatcttt
      421 gataaataaa attgccgtat tgtcaggctc tttattttta tcccaaccat tagccaaagt
      481 cgagttctca aaattgagct taaatttctc atcactatat ggcttttgag ttatatagtt
      541 tctaattttg ttataaagag gcactatatt cgctagctca aagtagcact cctcaaatac
      601 tagataaaaa tgctcatcct tgtctaaaat atttgcctta tcttctgact gactaatatg
      661 aaatattttt agtttatgta agagattatt agtttgatct aaaagatcct tgatagcttt
      721 aacatcatct tccgcactag cttgaagtag gtctttttta ccttgatttt gatatttgat
      781 agatatctgt gccaaattgt ctttgttttg agctatttca tcaaatatca tcggaatagc
      841 cgcaaagttt gcaagtattt cttcaaacct acactgttta tctatatctc tatgcttatt
      901 aaattcttct aaggcaagct ttatagtttc tagagataag tattttgctt tttcagtttt
      961 tttggctatt aattcttgct ctttcttact agggttatca agatttttag gtgctatttg
     1021 ttgagttata tattctagta ccgctgtacc aataacacta taatcatcaa aaacttgttg
     1081 tgatagatca gtaagagatt tatcattttt aaaataaatt ttactcaaat caagtttttg
     1141 agcttttaaa tcatcaaata ataaagatag tgtttcttta atagattttt cttctactgt
     1201 tttaaaagct gctatttgct cataaaaact ttgcatcgtt gtaactacat cactatcatc
     1261 ttctaactta tcaattacaa aagatttaga ttctgtatca cttaaaattt gcttaaataa
     1321 aacactcatt ttatattttt tgagtgtttt atcatttatt tgctgtgagt atagatttat
     1381 atattcattt ataccttttc tctttgtatt ttcaccattt acaaatttac caccaataat
     1441 agtattaaat ttagtaatac cactttgatt tagataatta ttaaagtttg ctatctcaaa
     1501 aacttcatca agtgaaaaaa ctctttgatt aacttcagat gttttgtagt caatatcaaa
     1561 ggttagctct tctgccaaat cttttttaat ttgttcatag tttatagctt ctggagcttt
     1621 gtcttttaaa ctctcatact tagctttatt ttctagaaat ttaggcaaat tatcatctac
     1681 tatcctataa ataatagatg taggaatatc attgctacta taaacatttt ttctattttc
     1741 atgaaaaccc ttaaaataag ttgtccaacc tttaaaagat ttgattattt ctaacgcctc
     1801 atctatatct gtgatatcac tattggcttt aaatagttct ataccattat ccttagattg
     1861 ctttagccat agaattaaat ctgactcttg ccctttttta gcatcgataa ggttttgatt
     1921 aaacaaattc ttaaatttct ctgagtcctt tatatattca gatatttgtt tctttatcgt
     1981 atcttttgca cttttaaaat ctttttgtag attatcatca tcactctttt taagtttaaa
     2041 ataaacatca gaatagtttt gtaataaatc ttcgctaata caaaccgaac ttaatatctc
     2101 ctctataaaa aactgatgat atttatcaat tatttgttta gcctttttgt agtctttagc
     2161 tcttttctca tcatctaaaa tcaaacctct tgcttttatg ttttcaagtg ttttaccctg
     2221 tgggattaac tcaaatctta gagttttact taaactatat ttattaacaa attcttgata
     2281 aattgacata gatcctttct cctctttttt attcgactat aacaaaccat tttcttgcgt
     2341 aaacctgtac gatcctacag gtagatcttt aatttttaaa gtatgggcaa tcaattgctc
     2401 ctgttaaaat tgctttagaa atactttggc agcggtttgt tgtattgagt ttcatttgcg
     2461 cattggttaa atggaaagtg acagtacgct cactgcagcc taatattttt gaaatatccc
     2521 aagagctttt tccttcgcat gcccacgcta aacattcttt ttctcttttg gttaaatcgt
     2581 tgtttgattt attatttgct atatttattt ttcgataatt atcaactaga gaaggaacaa
     2641 ttaatggtat gttcatacac gcatgtaaaa ataaactatc tatatagttg tctttttctg
     2701 aatgtgcaaa actaagcatt ccgaagccat tgttagccgt atgaataggg aaactaaacc
     2761 cagtgataag acctgatgtt ttcgcttctt taattacatt tggagatttt ttatttacag
     2821 cattgttttc aaatatattc caattaattg gtgaatgatt ggagttagaa taatctacta
     2881 taggatcata ttttattaaa ttagcgtcat cataatattg cctccatttt ttagggtaat
     2941 tatctagaat tgaaatatca gatttaacca tagaatgagg ataaatgatc gcgagtaaat
     3001 aatattcaca atgtaccatt ttagtcatat cagataagca ttgattaata tcattattgc
     3061 ttctacaagc tttaatttta ttaattattc tgtatgtgtc gtcggcattt atgtttttca
     3121 tacccatctc tttatcctta cctattgttt gtcgcaagtt ttgcgtgtta tatatcatta
     3181 aaacggtaat ggattgacat ttgattctaa taaattggat ttttgtcaca ctattgtatc
     3241 gctgggaata caattactta acataagcac ctgtaggatc gtacaggttt acgcaagaaa
     3301 atggtttgtt atagtcgaat aaacgcaagg gaggttggta tgactataat gataaaaaaa
     3361 tcggattttt tggcaattcc atcggaggag tataaaggta ttctaagtct tcgttatcaa
     3421 gtgtttaagc aaagacttga gtgggactta gttgtagaaa ataaccttga atcagatgag
     3481 tatgataact caaatgcaga atatatttat gcttgtgatg atactgaaaa tgtaagtgga
     3541 tgctggcgtt tattacctac aacaggtgat tatatgctga aaagtgtttt tcctgaattg
     3601 cttggtcaac agagtgctcc caaagatcct aatatagtcg aattaagtcg ttttgctgta
     3661 ggtaaaaata gctcaaagat aaataactct gctagtgaaa ttacaatgaa actatttgaa
     3721 gctatatata aacacgctgt tagtcaaggt attacagaat atgtaacagt aacatcaaca
     3781 gcaatagagc gatttttaaa gcgtattaaa gttccttgtc atcgtattgg agacaaagaa
     3841 attcatgtat taggtgatac taaatcggtt gtattgtcta tgcctattaa tgaacagttt
     3901 aaaaaagcag tcttaaatta atctagataa gttcaggatg aattctctag tccaaccttt
     3961 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     4021 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     4081 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     4141 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     4201 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     4261 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     4321 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     4381 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     4441 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     4501 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     4561 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     4621 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgt agttcattca gggcaccgga
     4681 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     4741 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     4801 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     4861 ctcttgatca aatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     4921 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc
     4981 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     5041 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     5101 cccaggtggc acttttcggg gaaatgtgcg cgcccgcgtt cctgctggcg ctgggcctgt
     5161 ttctggcgct ggacttcccg ctgttccgtc agcagctttt cgcccacggc cttgatgatc
     5221 gcggcggcct tggcctgcat atcccgattc aacggcccca gggcgtccag aacgggcttc
     5281 aggcgctccc gaaggtctcg ggccgtctct tgggcttgat cggccttctt gcgcatctca
     5341 cgcgctcctg cggcggcctg tagggcaggc tcatacccct gccgaaccgc ttttgtcagc
     5401 cggtcggcca cggcttccgg cgtctcaacg cgctttgaga ttcccagctt ttcggccaat
     5461 ccctgcggtg cataggcgcg tggctcgacc gcttgcgggc tgatggtgac gtggcccact
     5521 ggtggccgct ccagggcctc gtagaacgcc tgaatgcgcg tgtgacgtgc cttgctgccc
     5581 tcgatgcccc gttgcagccc tagatcggcc acagcagccg caaacgtggt ctggtcgcgg
     5641 gtcatctgcg ctttgttgcc gatgaactcc ttggccgaca gcctgccgtc ctgcgtcagc
     5701 ggcaccacga acgcggtcat gtgcgggctg gtttcgtcac ggtggatgct ggccgtcacg
     5761 atgcgatccg ccccgtactt gtccgccagc cacttgtgcg ccttctcgaa gaacgccgcc
     5821 tgctgttctt ggctggccga cttccaccat tccgggctgg ccgtcatgac gtactcgacc
     5881 gccaacacag cgtccttgcg ccgcttctct ggcagcaact cgcgcagtcg gcccatcgct
     5941 tcatcggtgc tgctggccgc ccagtgctcg ttctctggcg tcctgctggc gtcagcgttg
     6001 ggcgtctcgc gctcgcggta ggcgtgcttg agactggccg ccacgttgcc cattttcgcc
     6061 agcttcttgc atcgcatgat cgcgtatgcc gccatgcctg cccctccctt ttggtgtcca
     6121 accggctcga cgggggcagc gcaaggcggt gcctccggcg ggccactcaa tgcttgagta
     6181 tactcactag actttgcttc gcaaagtcgt gaccgcctac ggcggctgcg gcgccctacg
     6241 ggcttgctct ccgggcttcg ccctgcgcgg tcgctgcgct cccttgccag cccgtggata
     6301 tgtggacgat ggccgcgagc ggccaccggc tggctcgctt cgctcggccc gtggacaacc
     6361 ctgctggaca agctgatgga caggctgcgc cagcccacga gcttgaccac agggattgcc
     6421 caccggctac ccagccttcg accacatacc caccggctcc aactgcgcgg cctgcggcct
     6481 tgccccatca atttttttaa ttttctctgg ggaaaagcct ccggcctgcg gcctgcgcgc
     6541 ttcgcttgcc ggttggacac caagtggaag gcgggtcaag gctcgcgcag cgaccgcgca
     6601 gcggcttggc cttgacgcgc ctggaacgac ccaagcctat gcgagtgggg gcagtcgaag
     6661 ggcgaagccc gcccgcctgc cccccgagcc tcacggcggc gagtgcgggg gttccaaggg
     6721 ggcagcgcca ccttgggcaa ggccgaaggc cgcgcagtcg atcaacaagc cccggagggg
     6781 ccactttttg ccggaggggg agccgcgccg aaggcgtggg ggaaccccgc aggggtgccc
     6841 ttctttgggc accaaagaac tagatatagg gcgaaatgcg aaagacttaa aaatcaacaa
     6901 cttaaaaaag gggggtacgc aacagctcat tgcggcaccc cccgcaatag ctcattgcgt
     6961 aggttaaaga aaatctgtaa ttgactgcca cttttacgca acgcataatt gttgtcgcgc
     7021 tgccaaaaag ttgcagctga ttgcgcatgg tgccgcaacc gtgcggcacc cctaccgcat
     7081 ggagataagc atggccacgc agtccagaga aatcggcatt caagccaaga acaagcccgg
     7141 tcactgggtg caaacggaac gcaaagcgca tgaggcgtgg gccgggctta ttgcgaggaa
     7201 acccacggcg gcaatgctgc tgcatcacct cgtggcgcag atgggccacc agaacgccgt
     7261 ggtggtcagc cagaagacac tttccaagct catcggacgt tctttgcgga cggtccaata
     7321 cgcagtcaag gacttggtgg ccgagcgctg gatctccgtc gtgaagctca acggccccgg
     7381 caccgtgtcg gcctacgtgg tcaatgaccg cgtggcgtgg ggccagcccc gcgaccagtt
     7441 gcgcctgtcg gtgttcagtg ccgccgtggt ggttgatcac gacgaccagg acgaatcgct
     7501 gttggggcat ggcgacctgc gccgcatccc gaccctgtat ccgggcgagt agcaactacc
     7561 gaccggcccc ggcgaggagc cgcccagcca gcccggcatt ccgggcatgg aaccagacct
     7621 gccagccttg accgaaacgg aggaatggga acggcgcggg cagcagcgcc tgccgatgcc
     7681 cgatgagccg tgttttctgg acgatggcga gccgttggag ccgccgacac gggtcacgct
     7741 gccgcgccgg tagcacttgg gttgcgcagc aacccgtaag tgcgctgttc cagactatac
     7801 tagactgcag gtcctgaagt taacaaaaaa aatgaccttc ataaatcgct aatctacaac
     7861 agtagaaatt atttaaagtt cttagaccgg ctactaatac aaaaggctct agacatctac
     7921 aacagtagaa attatttaaa gttcttagac tgaattatct gaaggcacag gaatagtagc
     7981 atctacaaca gtagaaatta tttaaagttc ttagacgctc ctcgcccttg ctcaccatgc
     8041 tcgagccctg atctacaaca gtagaaatta tttaaagttc ttagacccgt ttttgcctaa
     8101 atcagcgcta gcattatacc taggactgag ctagctgtca actcgagtta gttattccta
     8161 ttctgcacga actcaaaata ctcttcattt ttgataacca aattgagttt tttgccctct
     8221 tgattatttt tgatcctacc tagtagcatc agacctttta gcccaatatg ataagcacca
     8281 ttggcatcag catcttgagg catatttttt ggcgcctgtc gcgaatcaaa gaaattgcca
     8341 tttacatctg ctactggtga aattagataa tctaactcag tacctgtttt tgagttacgc
     8401 atttgtaaga tagtatttag gacactagtt agcttagcaa aaaacttttt gtcgctctca
     8461 ccgcaaatag ctgctttgat acattcgcca tgcccatatt cgatagaata atcttttagc
     8521 aatttctcca actctttagt tggataaact tctcgagtat cccaattatg atttttatct
     8581 gaatttctaa agttaatcaa tctactccca aagctagcta tagtccactt gcctttggca
     8641 gccttgtcac caaagttttt ataatcaaaa ctaaactcaa aatagccctt atcaaggtta
     8701 taacaaatct tgtcaaactt actaaagaac tcttgagatt tgctgacact ttcatactta
     8761 ggatataact gatttacaaa accagttaca ggacaaattt ttgaagtaaa accagctggt
     8821 acatagtaga taatacctgt ttgtttaccc atctttttaa aagtctcaaa aggtgctgtt
     8881 agctgataag ctctaagcac tcccccagtt ttatcaaact cattatcttt gaaaactaga
     8941 tagtttagtt tctcaattag cattttttct aacttttgat agacctgctt ctctaccttg
     9001 aaacgccctc ttttaaatcc aaaatttaaa tcctcaaaaa ccacaatagc attatactct
     9061 ataactagct tagctatttc atgaactacc tgagatagat agccctcttt catctctttg
     9121 atgttattta tctttttcca gtctttccta gctgaatccc tatctttctc tattgcagca
     9181 agcttatcat ggtagtttgt tttcattcta tcattaccaa tgatgttgaa agtatcttgt
     9241 ttgatgatat tgcctttacc atctaccaaa gtatagtaag ctaaatgtct ttcacctcta
     9301 gctatactta atatatgaac atcatttgct ttttctttta gcaataaatt gatttcatca
     9361 ttaaacttat tagctccact agatttaaaa ttgattgtaa taggacagtg aaagaaaaac
     9421 ttatcttcag taaagcgttt atctttgatt aaatcatatt caaaaacact ctctttttta
     9481 ggattatctt tgtttttatt agctattgcc tctttagctg ggtgagtgat ttttttaggt
     9541 attgattgtt tacgataaaa aagctctgcc tcaccattta gcttataaac cacatcttga
     9601 agatttctct catcaaacag cgctttccaa tataaagtat gtagatttgg tcgccctttg
     9661 ctataagctg aaaaatcttt attatagatt tggaataggt acaatttacc ctgattaact
     9721 acgctatcaa tatagctctc tgatatattt tcaaaagtta g
//
