LOCUS       sequence-197167-        2936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Repress mCherry and GFP independently, with different strengths.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2936)
  AUTHORS   Vigouroux A, Oldewurtel E, Cui L, Bikard D, van Teeffelen S
  TITLE     Tuning dCas9's ability to block transcription enables robust, 
            noiseless knockdown of bacterial genes.
  JOURNAL   Mol Syst Biol. 2018 Mar 8;14(3):e7899.
  PUBMED    29519933
REFERENCE   2  (bases 1 to 2936)
  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: "Mol Syst 
            Biol."; date: "2018-03-8"; volume: "14(3)"; pages: "e7899"
COMMENT     Sequence Label: pCRRNA-G10-R0
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(93..179)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    209..340
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   341..376
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   407..442
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   473..508
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_RNA        629..707
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     primer_bind     complement(852..870)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1049..1843
                     /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(1103..1122)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1713..1732
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1874..1968
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2056..2644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2545..2564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtaacctg tcggatcacc ggaaaggacc cgtaaagtga taatgattat catctacata
       61 tcacaacgtg cgtggagggt ctagggcggc ggatttgtcc tactcaggag agcgttcacc
      121 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
      181 tgcctctagc acgcgtacca tgggatccta tttcttaata actaaaaata tggtataata
      241 ctcttaataa atgcagtaat acaggggctt ttcaagactg aagtctagct gagacaaata
      301 gtgcgattac gaaatttttt agacaaaaat agtctacgag gttttagagc tatgctgttt
      361 tgaatggtcc caaaaccctt caccttgtgg tgcttgagag aatttggttt tagagctatg
      421 ctgttttgaa tggtcccaaa acatcgcaca tcctggtcgc gacattaaga gtgttttaga
      481 gctatgctgt tttgaatggt cccaaaactt cagcacactg agacttgttg agttgaattc
      541 atgagtatta agtattgttt tatggctgat aaatttcttt gaatttctcc ttgattattt
      601 gttataaaag ttataaaata atcttgttgg aaccattcaa aacagcatag caagttaaaa
      661 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgatact
      721 tctattctac tctgactgca aaccaaaaaa acaagcgctt tcaaaacgct tgttttatca
      781 tttttaggga aattaatctc ttaatccttt tatcattcta catttaggcg ctgccatctt
      841 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
      901 ttcttagacg tcggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
      961 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
     1021 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     1081 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     1141 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     1201 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     1261 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     1321 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     1381 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     1441 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     1501 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     1561 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     1621 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     1681 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     1741 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     1801 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     1861 gagagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     1921 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagt
     1981 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2041 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2101 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2161 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2221 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2281 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2341 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2401 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2461 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2521 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2581 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2641 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2701 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2761 agctgatacc gctcgccgca gccgaacgcc ccaaaaagcc tcgctttcag cacctgtcgt
     2821 ttcctttctt ttcagagggt attttaaata aaaacattaa gttatgacga agaagaacgg
     2881 gaacgcctta aaccggaaaa ttttcataaa tagcgaaaac ccgcgaggtc gccgcc
//
