LOCUS       sequence-484256-       10543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Violacein biosynthesis pathway induced by CRISPRa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10543)
  AUTHORS   Stohr AM, Hansen H, Richards B, Park H, Goncalves AG, Agrawal A, 
            Blenner M, Chen W
  TITLE     Metabolite-responsive scaffold RNAs for dynamic CRISPR 
            transcriptional regulation.
  JOURNAL   Nucleic Acids Res. 2025 Nov 13;53(21):gkaf1290. doi: 
            10.1093/nar/gkaf1290.
  PUBMED    41277686
REFERENCE   2  (bases 1 to 10543)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2025-11-13"; volume: "53(21):gkaf1290. doi"; 
            pages: " 10.1093/nar/gkaf1290"
COMMENT     Sequence Label: pColE1-MP-Violacein
FEATURES             Location/Qualifiers
     source          1..10543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(19..607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(99..118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(778..1569,1570..1638))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1401..1420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1639..1743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1811..1829
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             2395..2406
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             3711..3733
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             6783..6794
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             7419..7430
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             8739..8750
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(10391..10408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
       61 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1381 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1741 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1801 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct ggcgaatcct
     1861 ctgaccagcc agaaaacgac ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca
     1921 gcaagtgggg gacagcagaa gacctgaccg ccgcagagtg gatgtttgac atggtgaaga
     1981 ctatcgcacc atcagccaga aaaccgaatt ttgctgggtg ggctaacgat atccgcctga
     2041 tgcgtgaacg tgacggacgt aaccaccgcg acatgtgtgt gctgttccgc tgggcatgcc
     2101 aggacaactt ctggtccggt aacgtgctga gcccggccaa gcttagcccg aaaagtgcca
     2161 cctgtggcaa ttccgacgtc agcatttgcg atcattcacg cagcgcttat tcagttgctc
     2221 actgcgatgt cataatcatc gctacgagct gtgaaagatg cataaagctc gtacgacgcg
     2281 ttcgctcgtc tcctcacttc tcctaactct cacacgtggc tgcaccggcc ccccccgctg
     2341 ccgcgggccg ttgacagttt cggggccgaa agggattgta cgagcgaatt cattaaagag
     2401 gagaaaggta ccatgttaaa tttagagcat tcagtttcaa ttgtcggagc tggaatttct
     2461 gggatagtgt gtgcaattac attagctcgt tcaagggcat tagaaggtta cgcaatagat
     2521 gtctatgaac aaagcccccg cgttggtgga cggatccaaa caaacttgtt gagcaatcat
     2581 tacatcgatt taggtgccgc tcgtttttgt tcacaattgc atggtaatgt atatgatcta
     2641 gtcaagaggt ttgaactcga atttgaaccg tttcctttta ctcaacttag atttactcag
     2701 ccaattcaaa aagaactcaa gactattttg tctaatctaa aaccgttaat cgatgaacat
     2761 aaagaagatt cttttgtcga atttgttacg agctatttgg gtaaagaaaa agcaaaagca
     2821 gtaatcttag cactgggcta cgactcactc tctatgccaa taatttctcc acatattgct
     2881 tacgatatca ttgaaaaaca cccagaaact caatgtttca ctgaaaattg tgggaatgaa
     2941 tggtttagtt tgaaagaggg attggcgtcg ttgatcaagg cgctatatga agaggcgtta
     3001 aaactcggga ttaagtttca ttttgaatct acattagttc aaataagcaa tcgaggtaac
     3061 gcccacagct tggtgattgc cgataatcaa aatcaacaaa aagagatcag taaagggctt
     3121 aagatcatta ctttgcctcc atctcgaatg gaaaaattag ataagcaatt tcctaaacgt
     3181 tggagcaact acaattatgg ttctagcccc ttgtttaaag gatttttctt ttactccgaa
     3241 ccgtggtggc gagagttagg gttgacggat cacgtcgtaa ttacccaaaa tccgatccgt
     3301 aaattgtatt tcaaagatga aaaatacgcg tttttctata ctgatagtga caacgcgtta
     3361 tattggcgag atgtgctttc tttaggcgaa gatcattacc ttacaacggt taaaaagctt
     3421 atcgcggagg cactaaactt acctcaagat gtcattccaa atccagagtt acacaaaaac
     3481 aaattttggc ttcatggtgt cgaattttca aaagaatgtt gccctaaaca tccattttat
     3541 caaacacttt ctgatgacag cgttatctct ttgtctgacg cctatacaca ccattgtgga
     3601 tggatagaag ggggcatctt agccggcaag tttacagcgg aatcgatcat cagccagttg
     3661 cctacacatc gtgaaactgt gaaagaagta gtataccttt gagaaataat tttgtttaac
     3721 tttaagaagg agatatacat atgagtgttt tagattttcc tcgaatacat tttaagggta
     3781 tggctcgggt taatatacca accgcaaacc gaaataccca aaacacattg gatacgacaa
     3841 caaataccgt gtttcaattc ggccagccgg ttgacattga cttgccgccg gcacgtttcc
     3901 acgaatacct ccactcactt ggtccaaaag ttaatcgctt tggtaaaaaa gatgaaaatg
     3961 gagaattcag tgtagcgtgt ggctacaaca tgcttggtaa taatcatttt tcatggcacg
     4021 acgctaaagt tacggctttg caattgcgcc cgggtgattg tgttgttcaa gatccactca
     4081 tagggtgttc tgttaattta tggggacact acaatgagta ccttagaacc tcattcaatc
     4141 gtgctcgctg ggttgataat gacccgaccc gaagtgactc ggcactcatc tatgctggac
     4201 aaattacatt aactgcaaat ggcagtgagg caagtgaaaa gcatgtgatg tctgctgaca
     4261 ttgattgtac acatggagtg agatggataa atcctagctt tattaatagc ctcgacgaac
     4321 atgcgtttgc tgaagatatt gcagcagccc gtttgtttca gtttgctgtc aataagaaca
     4381 gcaaggattt cttattgaat caattgggct tgcagtcaga atttattgaa caattacgaa
     4441 tagcactaga acaagaagat gttaagggct tagttgttca atattgtgta agcaatctat
     4501 cgcctcctgt agcgcccaat atgccggtct tttgtgattt acatggctgc atagggcttt
     4561 ggcgagaaag tgaaatggcc acttgtcctg caggtcggat tttacagcca gaatgcccac
     4621 agcaatttgc accgattaca ttaaaacagg cagggcaatg gttgagtata aatatgccga
     4681 ttagtatcgg tcatactcat cacttagatc cattacctgt attatctgga atgcctcctc
     4741 gattagcagc aaaaacgccc ttgggtgtct tgcaactcaa gacggccagt gatgagttaa
     4801 tcgccgagat cccagaggtg ctttataacg cgtgtttgga tacaggaggg atcatcgatg
     4861 tgacaataca aacccataca gacggacttg aattacagtc attaaagctg gtaagccagt
     4921 ctcattcgtg gcatgaaatc gattggcaag ttcaagccga acaacacatc atatatatgg
     4981 aatctccaaa cctaatcgag ttgcaaaaaa gcaagcaata tattgatgta tatagctatt
     5041 ttcgtggaat tcttaagcca atcaacggtt taacaagtta taacgcggga ccacaacgag
     5101 tgcaatgtga cgagttcatt gaaacgaatg aacaaggacg tggtttgttt ctagttgaag
     5161 cgttaacacc agggagtggt gaagtttttc taggagaaca atatagtcgt gtccagatcc
     5221 gtgtattaag taatgactgg gcgttactta atgtccctga tgaacaggtt gactatgact
     5281 ttctttataa aaatgtcatg gcctactatg agtttgttta tccgttcatg gcagacaaag
     5341 tgtttagcat ggctgataaa tgtaaatgcg aaacgtatgc acgcttaatg tggcaaatgt
     5401 gcgatcccaa aaataaagag aaaagctact acatgcctag tacgcgggag atgtcacaag
     5461 cacaatcgtt tctatttttg aaatatttga gtaacgtaga aaagtcggca atccccaagg
     5521 ttttgcctta ttgcaataca gacgttacgg ttacaggcaa aataaaaaac cgtacacaat
     5581 taattcgggc attaaaagac tctgtcgacc tagagttgtc gattatgctg cagtatttgt
     5641 atagcgccta ttcactacca aattacgccg ccggagaggc gttggtaaac gcaaaaagat
     5701 ggactcaaga acaacttgaa ttagtgaatg gtggcgcaga taggcggcac aacagcggtt
     5761 ggcgaggtgc tattcttgaa gttgctcacg aagagatgat ccattatttg gtcgtcaata
     5821 atttgcttat gtccttaggc gaaccttttt actgtggcaa cccggtactt ggccaacaag
     5881 cgaacgaaaa atttggactc gatacggatt tttcttacga gccctttggc ttacagtcca
     5941 tcacgaaatt tgtacgcttt gagtggccac gctttattcc gaccataggt ctttcaattg
     6001 ctgatttcta tgccgacatt aagcgtgcct ttgagcaatt accagacctc tatcctaaag
     6061 atgcgaaggg ttcgggggga gagcatcact tgtttttgaa tgaagtgatt aaccgtcaat
     6121 acccggcata ccaattcgaa gtgtataacc gacagactgc tttgtttgca attgaatttg
     6181 ttacagagca aggcgaaggt gtgactgcac agtcagaaca attcgaatta ggtcatttcc
     6241 accgcttaag acagatagcc atggcattaa ctaatcaaga acagcctttt gaaccggcgt
     6301 atcccgtgct taaaaaccct gtttttaaac ctagagcggg atgtaattta gtcaccgacc
     6361 cagaatcaca agccttaatg gcgttttatc aacagtgtca tgaattgatg tttcaaatca
     6421 tgatgcatca tttttcgcaa cgacctctca tcagcatgcg gcgctcgcga ttgatgaatg
     6481 ccgccatcga cctgatgaca ggaatattac gtccattgtc tgtgaaatta atgacgatgc
     6541 cgtctgggac taaaggtcga aatgccggtc cgccagtgcc agagtcggtc agttctttcg
     6601 tcgataagga ttatgctggc ggttgccaac aattgtcatg gcgctgcgca cagttggcgg
     6661 cagcggcaaa agcgttaaac ctaaatgtaa ctacacaaat cgaattattg gagtttttcc
     6721 aacaacaact ggctgacctc gcagccggaa ctctttcaaa ggaaggttag caattgtcta
     6781 gaaaagagga gaaatactac atggaattac gtaaagtaga tagagttccg ccaatgttac
     6841 ctgagtgttg gagcagtagt tatatttctt attggcttcc tatgaagcca gaagacgata
     6901 ttacttctgg gtattgttgg ttcgattatg ctcaagaggt ttgtcgtata gacggtttgt
     6961 ttaacccttg gagtgaaaca aaaatgggac accgtttgtg gatgtccgaa attatgtatc
     7021 cacgcagtta tgaatcgttt aaatctaaaa ttgcctatgc acgaacacat cgggataaac
     7081 cttcagcgtt taaagaaacg gtgttaaatg atgaggtaga cccatgccat gagcttattt
     7141 taacgcgtga tgtgttggta aaaaaccacg ctaagtatgc aggtgaacac accattttgg
     7201 gttataaagc ggatgcgtgg tgctttgaac gcccaaatga caaaggtcct gcaacgtatt
     7261 tttttaaaga agggacagat cagttgctcc gaatggtcac gggtgatccg aacgttcacg
     7321 cctctgtaag agactttcct aattttaata cgctgccgat acctaacacg atttttgatg
     7381 taacaagtct ctctcatagg aattgacaat tgtctagaaa agaggagaaa tactacatga
     7441 gtaaaataat tattgttggt ggtgggttgg caggcagcct gacggcaatc tatctagcga
     7501 aacaagggca cgaagttcac attatagaaa aacgtggaaa tcctttagaa agtcaggcgg
     7561 attatattga tcaagttagc gcgcgtgcga ttggtgtcag tatgacagta cgaggaattg
     7621 aagccgtttt agcagccggt attcccttgg acgaattgca ggcgtgcggc gttgaagttt
     7681 caggtatggc gttttttatc aaaggtcaac ctaaaatacg ggatttacca ccgttagata
     7741 cgttaaaacc gttgtcgtta agccgaagca tgtttcaggt attgcttaac aagtacgctc
     7801 ataaagcaaa agtacactat cactatctac aacgctgtgt cgacgttgat attgaaagag
     7861 cgtcagtgat tacacaaaac cagtatggcg aatttgttaa ccaccaaggt gatttgctga
     7921 ttgctgcgga tggcgcccgt tcagttgtcc gagacgcatt tcaaactcat agtcgtcgtt
     7981 ttgagtttca tcaatccttt tttaagcatg gttataaaac actcgtgatc ccagatgcca
     8041 cagaagtcgg tcttcgcaaa gacattttat atttctttgg tatggattct ggaggggaat
     8101 ttgcaggtcg tgcggcaact atccctgatg gcagtatcag ttttgctgtg tgtttacctt
     8161 ataccggaaa agtaagttta cactctgaag acacgcaagc aatgaaggat ttctttgaac
     8221 gctattacgc catgatacct aagcacattc gagagtcgat gtaccaacaa tttatggaaa
     8281 agccaagtaa tgacttaatt aatgttcgat cgtcaacctt tcattataaa aacagagcat
     8341 tgttgttagg ggatgcggcc catgctaccg cacctttttt aggacaggga atgaatatgg
     8401 ctttagaaga cgcctttgtc cttgatagat tgtttgccga gtttgaagga aacttaaccg
     8461 aaatcttgcc tcgttttact gacctacgca aaaaagaagc tgatgcgatg caagatatgg
     8521 cgcgggcaaa ctatgatgtc ttgtcttgct caaatccagt cttttttatg cgtgcgaaat
     8581 ttgcccgtta tatgacaaaa caattcccag atcgctatcc acccgatatg gctgaaaaat
     8641 tatactttac gtctatggcg tatagcgagc ttcgggatgt tcaacaaaaa cagaatgttt
     8701 ggtacaaaat agggagaatg aattaacaat tgtctagaaa agaggagaaa tactacatga
     8761 acattttagt gatcggggca ggtcctgccg gattaatgtt tgctagtcaa attaaaaaac
     8821 tgaatccaga ttgggatatt tgtattaaag aaaagaataa tcaagacgaa accgtaggtt
     8881 ggggcgtagt gctgccaggg cgcgcgccac atcatcccgc caatccactt tcgtatcttc
     8941 ctaattatga atccttggat gcacaatatt tagatgcctt taaaatgact caccacaatg
     9001 aaagtgcgtt aaaaagtaca gggctaacct tatgcggcgc tgagcgaaaa tcgatgatta
     9061 aggctatgcg agatttgtgt attcgtttgg gtatccatat tgattatgag caaccagttg
     9121 aaaatcttga attagctagc actcaatatg acttggttgt ggtatcaaac gggataaacc
     9181 acgccaacca attctatcgt gaagcgctaa aaccgaacat tgaatatggc acgaatcgct
     9241 acatgtggta tggcacgcct aatcttttta atgaaatgaa cctgattttt aaagcgcatt
     9301 ctaaaggtgt ttttgtcgct catgcctata aatattcgag tgacatgagt acttttgttg
     9361 ttgaatgtag cgaagagacg tatctaaagt caggtctaga cgaaatgtct gcagaccaag
     9421 cagaagcgtt tattgcatcc gtatttgaat ctgatttgaa cggaaagcct gttattgctc
     9481 aatcaggatt gaaatggcgt aatttcatga cgttaagtca tgaaaaggcc tatgatggaa
     9541 atattgtgtt attgggcgat gcgttgcaat cgggtcattt ttcaatcggg catggcacga
     9601 ccatggcagt ggttggcgca cagttactga ttaaatctgt ccatcaacat gcaaataacg
     9661 tcattgctgt ggtagagcac tttaatacgc gagtgatgcc tgtcatgaaa ttgtttgatc
     9721 aacatgcaca aacaagtcgc ctttggtttg aaagtgtagg agagctaatg catctaagta
     9781 cttctcaaat cgctcaaagt tttactgaac gtcgtgagca gctgccttct ttaccagaag
     9841 cgctagcaaa ggcattaggt tatgcgctgc aacgttaagg atccaaactc gagtaaggat
     9901 ctgtgctttt tttaacgcat gagaaagccc ccggaagatc accttccggg ggctttttta
     9961 ttgcgcggat ccttactccg tcgacaagct tgcggccgca ctcgaggaat tccgactcaa
    10021 ttagttcagt cagtttcagg atattagtca tctctacatt gattatgagt attcagaaat
    10081 tccttaaata ttctgacaaa tgctctttcc ctaaactccc cccataaaaa aacccgccga
    10141 agcgggtttt tacgttattt gcggattaac gattactcgt tatcagaacc gcccagacct
    10201 gcgttcagca gttctgccag gctggcagat gcgtcttccg gaattgatcc gtcgaccgat
    10261 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
    10321 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
    10381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    10441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    10501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaa
//
