LOCUS       sequence-205489-        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-G11-R0
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..32)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             211..1005
                     /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(265..284)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     875..894
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1036..1130
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1218..1806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1707..1726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2191..2277)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    2307..2438
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   2439..2474
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   2505..2540
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   2571..2606
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_RNA        2727..2805
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
ORIGIN
        1 ttaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
       61 gtttcttaga cgtcggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
      121 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct
      181 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
      241 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
      301 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
      361 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
      421 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
      481 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
      541 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
      601 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
      661 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
      721 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
      781 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
      841 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
      901 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
      961 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     1021 tcgagagctc gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     1081 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta
     1141 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     1381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1921 tgagctgata ccgctcgccg cagccgaacg ccccaaaaag cctcgctttc agcacctgtc
     1981 gtttcctttc ttttcagagg gtattttaaa taaaaacatt aagttatgac gaagaagaac
     2041 gggaacgcct taaaccggaa aattttcata aatagcgaaa acccgcgagg tcgccgcccc
     2101 gtaacctgtc ggatcaccgg aaaggacccg taaagtgata atgattatca tctacatatc
     2161 acaacgtgcg tggagggtct agggcggcgg atttgtccta ctcaggagag cgttcaccga
     2221 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     2281 cctctagcac gcgtaccatg ggatcctatt tcttaataac taaaaatatg gtataatact
     2341 cttaataaat gcagtaatac aggggctttt caagactgaa gtctagctga gacaaatagt
     2401 gcgattacga aattttttag acaaaaatag tctacgaggt tttagagcta tgctgttttg
     2461 aatggtccca aaacccttca ccttgtggtg cttcagagaa tttggtttta gagctatgct
     2521 gttttgaatg gtcccaaaac atcgcacatc ctggtcgcga cattaagagt gttttagagc
     2581 tatgctgttt tgaatggtcc caaaacttca gcacactgag acttgttgag ttgaattcat
     2641 gagtattaag tattgtttta tggctgataa atttctttga atttctcctt gattatttgt
     2701 tataaaagtt ataaaataat cttgttggaa ccattcaaaa cagcatagca agttaaaata
     2761 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgatacttc
     2821 tattctactc tgactgcaaa ccaaaaaaac aagcgctttc aaaacgcttg ttttatcatt
     2881 tttagggaaa ttaatctctt aatcctttta tcattctaca tttaggcgct gccatc
//
