LOCUS       sequence-469924-        9747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid pQdCas12a.sggfp(B) expresses the dCas12a endonuclease 
            and the sgRNA (design B) targeting the gfp gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9747)
  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 9747)
  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(B)
FEATURES             Location/Qualifiers
     source          1..9747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1756..1767
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        5683..5717
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(6057..6719)
                     /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"
     rep_origin      6720..7491
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             join(8971..9747,1..18)
                     /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(9025..9044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9635..9654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
       61 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg gactagagaa
      121 ttcatcctga acttatctag attaatttaa gactgctttt ttaaactgtt cattaatagg
      181 catagacaat acaaccgatt tagtatcacc taatacatga atttctttgt ctccaatacg
      241 atgacaagga actttaatac gctttaaaaa tcgctctatt gctgttgatg ttactgttac
      301 atattctgta ataccttgac taacagcgtg tttatatata gcttcaaata gtttcattgt
      361 aatttcacta gcagagttat ttatctttga gctattttta cctacagcaa aacgacttaa
      421 ttcgactata ttaggatctt tgggagcact ctgttgacca agcaattcag gaaaaacact
      481 tttcagcata taatcacctg ttgtaggtaa taaacgccag catccactta cattttcagt
      541 atcatcacaa gcataaatat attctgcatt tgagttatca tactcatctg attcaaggtt
      601 attttctaca actaagtccc actcaagtct ttgcttaaac acttgataac gaagacttag
      661 aataccttta tactcctccg atggaattgc caaaaaatcc gattttttta tcattatagt
      721 cataccaacc tcccttgcgt ttattcgact ataacaaacc attttcttgc gtaaacctgt
      781 acgatcctac aggtgcttat gttaagtaat tgtattccca gcgatacaat agtgtgacaa
      841 aaatccaatt tattagaatc aaatgtcaat ccattaccgt tttaatgata tataacacgc
      901 aaaacttgcg acaaacaata ggtaaggata aagagatggg tatgaaaaac ataaatgccg
      961 acgacacata cagaataatt aataaaatta aagcttgtag aagcaataat gatattaatc
     1021 aatgcttatc tgatatgact aaaatggtac attgtgaata ttatttactc gcgatcattt
     1081 atcctcattc tatggttaaa tctgatattt caattctaga taattaccct aaaaaatgga
     1141 ggcaatatta tgatgacgct aatttaataa aatatgatcc tatagtagat tattctaact
     1201 ccaatcattc accaattaat tggaatatat ttgaaaacaa tgctgtaaat aaaaaatctc
     1261 caaatgtaat taaagaagcg aaaacatcag gtcttatcac tgggtttagt ttccctattc
     1321 atacggctaa caatggcttc ggaatgctta gttttgcaca ttcagaaaaa gacaactata
     1381 tagatagttt atttttacat gcgtgtatga acataccatt aattgttcct tctctagttg
     1441 ataattatcg aaaaataaat atagcaaata ataaatcaaa caacgattta accaaaagag
     1501 aaaaagaatg tttagcgtgg gcatgcgaag gaaaaagctc ttgggatatt tcaaaaatat
     1561 taggctgcag tgagcgtact gtcactttcc atttaaccaa tgcgcaaatg aaactcaata
     1621 caacaaaccg ctgccaaagt atttctaaag caattttaac aggagcaatt gattgcccat
     1681 actttaaaaa ttaaagatct acctgtagga tcgtacaggt ttacgcaaga aaatggtttg
     1741 ttatagtcga ataaaaaaga ggagaaagga tctatgtcaa tttatcaaga atttgttaat
     1801 aaatatagtt taagtaaaac tctaagattt gagttaatcc cacagggtaa aacacttgaa
     1861 aacataaaag caagaggttt gattttagat gatgagaaaa gagctaaaga ctacaaaaag
     1921 gctaaacaaa taattgataa atatcatcag ttttttatag aggagatatt aagttcggtt
     1981 tgtattagcg aagatttatt acaaaactat tctgatgttt attttaaact taaaaagagt
     2041 gatgatgata atctacaaaa agattttaaa agtgcaaaag atacgataaa gaaacaaata
     2101 tctgaatata taaaggactc agagaaattt aagaatttgt ttaatcaaaa ccttatcgat
     2161 gctaaaaaag ggcaagagtc agatttaatt ctatggctaa agcaatctaa ggataatggt
     2221 atagaactat ttaaagccaa tagtgatatc acagatatag atgaggcgtt agaaataatc
     2281 aaatctttta aaggttggac aacttatttt aagggttttc atgaaaatag aaaaaatgtt
     2341 tatagtagca atgatattcc tacatctatt atttatagga tagtagatga taatttgcct
     2401 aaatttctag aaaataaagc taagtatgag agtttaaaag acaaagctcc agaagctata
     2461 aactatgaac aaattaaaaa agatttggca gaagagctaa cctttgatat tgactacaaa
     2521 acatctgaag ttaatcaaag agttttttca cttgatgaag tttttgagat agcaaacttt
     2581 aataattatc taaatcaaag tggtattact aaatttaata ctattattgg tggtaaattt
     2641 gtaaatggtg aaaatacaaa gagaaaaggt ataaatgaat atataaatct atactcacag
     2701 caaataaatg ataaaacact caaaaaatat aaaatgagtg ttttatttaa gcaaatttta
     2761 agtgatacag aatctaaatc ttttgtaatt gataagttag aagatgatag tgatgtagtt
     2821 acaacgatgc aaagttttta tgagcaaata gcagctttta aaacagtaga agaaaaatct
     2881 attaaagaaa cactatcttt attatttgat gatttaaaag ctcaaaaact tgatttgagt
     2941 aaaatttatt ttaaaaatga taaatctctt actgatctat cacaacaagt ttttgatgat
     3001 tatagtgtta ttggtacagc ggtactagaa tatataactc aacaaatagc acctaaaaat
     3061 cttgataacc ctagtaagaa agagcaagaa ttaatagcca aaaaaactga aaaagcaaaa
     3121 tacttatctc tagaaactat aaagcttgcc ttagaagaat ttaataagca tagagatata
     3181 gataaacagt gtaggtttga agaaatactt gcaaactttg cggctattcc gatgatattt
     3241 gatgaaatag ctcaaaacaa agacaatttg gcacagatat ctatcaaata tcaaaatcaa
     3301 ggtaaaaaag acctacttca agctagtgcg gaagatgatg ttaaagctat caaggatctt
     3361 ttagatcaaa ctaataatct cttacataaa ctaaaaatat ttcatattag tcagtcagaa
     3421 gataaggcaa atattttaga caaggatgag catttttatc tagtatttga ggagtgctac
     3481 tttgagctag cgaatatagt gcctctttat aacaaaatta gaaactatat aactcaaaag
     3541 ccatatagtg atgagaaatt taagctcaat tttgagaact cgactttggc taatggttgg
     3601 gataaaaata aagagcctga caatacggca attttattta tcaaagatga taaatattat
     3661 ctgggtgtga tgaataagaa aaataacaaa atatttgatg ataaagctat caaagaaaat
     3721 aaaggcgagg gttataaaaa aattgtttat aaacttttac ctggcgcaaa taaaatgtta
     3781 cctaaggttt tcttttctgc taaatctata aaattttata atcctagtga agatatactt
     3841 agaataagaa atcattccac acatacaaaa aatggtagtc ctcaaaaagg atatgaaaaa
     3901 tttgagttta atattgaaga ttgccgaaaa tttatagatt tttataaaca gtctataagt
     3961 aagcatccgg agtggaaaga ttttggattt agattttctg atactcaaag atataattct
     4021 atagatgaat tttatagaga agttgaaaat caaggctaca aactaacttt tgaaaatata
     4081 tcagagagct atattgatag cgtagttaat cagggtaaat tgtacctatt ccaaatctat
     4141 aataaagatt tttcagctta tagcaaaggg cgaccaaatc tacatacttt atattggaaa
     4201 gcgctgtttg atgagagaaa tcttcaagat gtggtttata agctaaatgg tgaggcagag
     4261 cttttttatc gtaaacaatc aatacctaaa aaaatcactc acccagctaa agaggcaata
     4321 gctaataaaa acaaagataa tcctaaaaaa gagagtgttt ttgaatatga tttaatcaaa
     4381 gataaacgct ttactgaaga taagtttttc tttcactgtc ctattacaat caattttaaa
     4441 tctagtggag ctaataagtt taatgatgaa atcaatttat tgctaaaaga aaaagcaaat
     4501 gatgttcata tattaagtat agctagaggt gaaagacatt tagcttacta tactttggta
     4561 gatggtaaag gcaatatcat caaacaagat actttcaaca tcattggtaa tgatagaatg
     4621 aaaacaaact accatgataa gcttgctgca atagagaaag atagggattc agctaggaaa
     4681 gactggaaaa agataaataa catcaaagag atgaaagagg gctatctatc tcaggtagtt
     4741 catgaaatag ctaagctagt tatagagtat aatgctattg tggtttttga ggatttaaat
     4801 tttggattta aaagagggcg tttcaaggta gagaagcagg tctatcaaaa gttagaaaaa
     4861 atgctaattg agaaactaaa ctatctagtt ttcaaagata atgagtttga taaaactggg
     4921 ggagtgctta gagcttatca gctaacagca ccttttgaga cttttaaaaa gatgggtaaa
     4981 caaacaggta ttatctacta tgtaccagct ggttttactt caaaaatttg tcctgtaact
     5041 ggttttgtaa atcagttata tcctaagtat gaaagtgtca gcaaatctca agagttcttt
     5101 agtaagtttg acaagatttg ttataacctt gataagggct attttgagtt tagttttgat
     5161 tataaaaact ttggtgacaa ggctgccaaa ggcaagtgga ctatagctag ctttgggagt
     5221 agattgatta actttagaaa ttcagataaa aatcataatt gggatactcg agaagtttat
     5281 ccaactaaag agttggagaa attgctaaaa gattattcta tcgaatatgg gcatggcgaa
     5341 tgtatcaaag cagctatttg cggtgagagc gacaaaaagt tttttgctaa gctaactagt
     5401 gtcctaaata ctatcttaca aatgcgtaac tcaaaaacag gtactgagtt agattatcta
     5461 atttcaccag tagcagatgt aaatggcaat ttctttgatt cgcgacaggc gccaaaaaat
     5521 atgcctcaag atgctgatgc caatggtgct tatcatattg ggctaaaagg tctgatgcta
     5581 ctaggtagga tcaaaaataa tcaagagggc aaaaaactca atttggttat caaaaatgaa
     5641 gagtattttg agttcgtgca gaataggaat aactaactcg agttgacagc tagctcagtc
     5701 ctaggtataa tgctagcgct gatttaggca aaaacgggtc taagaacttt aaataatttc
     5761 tactgttgta gatcagggct cgagcatggt gaggtctaag aactttaaat aatttctact
     5821 gttgtagatg ctactattcc tgtgccttca gataattcag tctaagaact ttaaataatt
     5881 tctactgttg tagatgtcta gagccttttg tattagtagc cggtctaaga actttaaata
     5941 atttctactg ttgtagatta gcgatttatg aaggtcattt tttttgttaa cttcaggacc
     6001 tgcagtctag tatagtctgg aacagcgcac ttacgggttg ctgcgcaacc caagtgctac
     6061 cggcgcggca gcgtgacccg tgtcggcggc tccaacggct cgccatcgtc cagaaaacac
     6121 ggctcatcgg gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc
     6181 aaggctggca ggtctggttc catgcccgga atgccgggct ggctgggcgg ctcctcgccg
     6241 gggccggtcg gtagttgcta ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca
     6301 tgccccaaca gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc actgaacacc
     6361 gacaggcgca actggtcgcg gggctggccc cacgccacgc ggtcattgac cacgtaggcc
     6421 gacacggtgc cggggccgtt gagcttcacg acggagatcc agcgctcggc caccaagtcc
     6481 ttgactgcgt attggaccgt ccgcaaagaa cgtccgatga gcttggaaag tgtcttctgg
     6541 ctgaccacca cggcgttctg gtggcccatc tgcgccacga ggtgatgcag cagcattgcc
     6601 gccgtgggtt tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc
     6661 acccagtgac cgggcttgtt cttggcttga atgccgattt ctctggactg cgtggccatg
     6721 cttatctcca tgcggtaggg gtgccgcacg gttgcggcac catgcgcaat cagctgcaac
     6781 tttttggcag cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat tacagatttt
     6841 ctttaaccta cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg cgtacccccc
     6901 ttttttaagt tgttgatttt taagtctttc gcatttcgcc ctatatctag ttctttggtg
     6961 cccaaagaag ggcacccctg cggggttccc ccacgccttc ggcgcggctc cccctccggc
     7021 aaaaagtggc ccctccgggg cttgttgatc gactgcgcgg ccttcggcct tgcccaaggt
     7081 ggcgctgccc ccttggaacc cccgcactcg ccgccgtgag gctcgggggg caggcgggcg
     7141 ggcttcgccc ttcgactgcc cccactcgca taggcttggg tcgttccagg cgcgtcaagg
     7201 ccaagccgct gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg tgtccaaccg
     7261 gcaagcgaag cgcgcaggcc gcaggccgga ggcttttccc cagagaaaat taaaaaaatt
     7321 gatggggcaa ggccgcaggc cgcgcagttg gagccggtgg gtatgtggtc gaaggctggg
     7381 tagccggtgg gcaatccctg tggtcaagct cgtgggctgg cgcagcctgt ccatcagctt
     7441 gtccagcagg gttgtccacg ggccgagcga agcgagccag ccggtggccg ctcgcggcca
     7501 tcgtccacat atccacgggc tggcaaggga gcgcagcgac cgcgcagggc gaagcccgga
     7561 gagcaagccc gtagggcgcc gcagccgccg taggcggtca cgactttgcg aagcaaagtc
     7621 tagtgagtat actcaagcat tgagtggccc gccggaggca ccgccttgcg ctgcccccgt
     7681 cgagccggtt ggacaccaaa agggaggggc aggcatggcg gcatacgcga tcatgcgatg
     7741 caagaagctg gcgaaaatgg gcaacgtggc ggccagtctc aagcacgcct accgcgagcg
     7801 cgagacgccc aacgctgacg ccagcaggac gccagagaac gagcactggg cggccagcag
     7861 caccgatgaa gcgatgggcc gactgcgcga gttgctgcca gagaagcggc gcaaggacgc
     7921 tgtgttggcg gtcgagtacg tcatgacggc cagcccggaa tggtggaagt cggccagcca
     7981 agaacagcag gcggcgttct tcgagaaggc gcacaagtgg ctggcggaca agtacggggc
     8041 ggatcgcatc gtgacggcca gcatccaccg tgacgaaacc agcccgcaca tgaccgcgtt
     8101 cgtggtgccg ctgacgcagg acggcaggct gtcggccaag gagttcatcg gcaacaaagc
     8161 gcagatgacc cgcgaccaga ccacgtttgc ggctgctgtg gccgatctag ggctgcaacg
     8221 gggcatcgag ggcagcaagg cacgtcacac gcgcattcag gcgttctacg aggccctgga
     8281 gcggccacca gtgggccacg tcaccatcag cccgcaagcg gtcgagccac gcgcctatgc
     8341 accgcaggga ttggccgaaa agctgggaat ctcaaagcgc gttgagacgc cggaagccgt
     8401 ggccgaccgg ctgacaaaag cggttcggca ggggtatgag cctgccctac aggccgccgc
     8461 aggagcgcgt gagatgcgca agaaggccga tcaagcccaa gagacggccc gagaccttcg
     8521 ggagcgcctg aagcccgttc tggacgccct ggggccgttg aatcgggata tgcaggccaa
     8581 ggccgccgcg atcatcaagg ccgtgggcga aaagctgctg acggaacagc gggaagtcca
     8641 gcgccagaaa caggcccagc gccagcagga acgcgggcgc gcacatttcc ccgaaaagtg
     8701 ccacctggga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag
     8761 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg
     8821 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     8881 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagattt
     8941 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     9001 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     9061 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     9121 accgacctgt ccggtgccct gaatgaacta caggacgagg cagcgcggct atcgtggctg
     9181 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     9241 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     9301 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     9361 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     9421 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     9481 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     9541 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     9601 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     9661 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     9721 tcgcagcgca tcgccttcta tcgcctt
//
