LOCUS       sequence-198936-        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-R11
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..33)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             212..1006
                     /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(266..285)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     876..895
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1037..1131
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1219..1807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1708..1727
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2192..2278)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    2308..2439
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   2440..2475
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   2506..2541
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   2572..2607
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_RNA        2728..2806
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
ORIGIN
        1 cttaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
       61 ggtttcttag acgtcggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
      121 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
      181 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
      241 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
      301 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
      361 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
      421 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
      481 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
      541 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
      601 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
      661 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
      721 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
      781 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
      841 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
      901 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
      961 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     1021 ttcgagagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     1081 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact
     1141 agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1321 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1861 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1921 gtgagctgat accgctcgcc gcagccgaac gccccaaaaa gcctcgcttt cagcacctgt
     1981 cgtttccttt cttttcagag ggtattttaa ataaaaacat taagttatga cgaagaagaa
     2041 cgggaacgcc ttaaaccgga aaattttcat aaatagcgaa aacccgcgag gtcgccgccc
     2101 cgtaacctgt cggatcaccg gaaaggaccc gtaaagtgat aatgattatc atctacatat
     2161 cacaacgtgc gtggagggtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg
     2221 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     2281 gcctctagca cgcgtaccat gggatcctat ttcttaataa ctaaaaatat ggtataatac
     2341 tcttaataaa tgcagtaata caggggcttt tcaagactga agtctagctg agacaaatag
     2401 tgcgattacg aaatttttta gacaaaaata gtctacgagg ttttagagct atgctgtttt
     2461 gaatggtccc aaaacccttc accttgtggt gcttgagaga atttggtttt agagctatgc
     2521 tgttttgaat ggtcccaaaa cagtttggcg gagacccacg cttcatacgg agttttagag
     2581 ctatgctgtt ttgaatggtc ccaaaacttc agcacactga gacttgttga gttgaattca
     2641 tgagtattaa gtattgtttt atggctgata aatttctttg aatttctcct tgattatttg
     2701 ttataaaagt tataaaataa tcttgttgga accattcaaa acagcatagc aagttaaaat
     2761 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttgatactt
     2821 ctattctact ctgactgcaa accaaaaaaa caagcgcttt caaaacgctt gttttatcat
     2881 ttttagggaa attaatctct taatcctttt atcattctac atttaggcgc tgccat
//
