LOCUS       sequence-194131-        3399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMS2-CYFP. Expresses MS2 protein-CYFP fusion in E. coli for TriFC 
            assay (kanR)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3399)
  AUTHORS   Gelderman G, Sivakumar A, Lipp S, Contreras L
  TITLE     Adaptation of Tri-molecular fluorescence complementation allows 
            assaying of regulatory Csr RNA-protein interactions in bacteria.
  JOURNAL   Biotechnol Bioeng. 2015 Feb;112(2):365-75. doi: 10.1002/bit.25351. 
            Epub 2014 Sep 26.
  PUBMED    25080893
REFERENCE   2  (bases 1 to 3399)
  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: "10.1002/bit.25351";
            journalName: "Biotechnol Bioeng"; date: "2015-02"; volume: "112"; 
            issue: "2"; pages: "365-75"
COMMENT     Sequence Label: pCML 221
FEATURES             Location/Qualifiers
     source          1..3399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     252..271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     468..485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    722..743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(758..763,764..781,782..788)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    796..812
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     801..823
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     820..836
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     820..836
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1231..1482
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1419..1440
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1489..1518
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1556..1575)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1675..1697
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2015..2809)
                     /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(2126..2145)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2736..2755
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      join(3205..3399,1..351)
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
ORIGIN
        1 atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa
       61 gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt
      121 ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc
      181 ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg
      241 gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga
      301 tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt
      361 gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc
      421 cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga
      481 gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt
      541 ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag
      601 tatatacact ccgctagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      661 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
      721 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
      781 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
      841 attacgccaa gcttgatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga
      901 actggcgacg tgactgtcgc cccaagcaac ttcgctaacg ggatcgctga atggatcagc
      961 tctaactcgc gttcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag
     1021 aatcgcaaat acaccatcaa agtcgaggtg cctaaaggcg cctggcgttc gtacttaaat
     1081 atggaactaa ccattccaat tttcgccacg aattccgact gcgagcttat tgttaaggca
     1141 atgcaaggtc tcctaaaaga tggaaacccg attccctcag caatcgcagc aaactccggc
     1201 atctacgcga aacagaaagt tatgaatcat gacaagcaga agaacggcat caaggtgaac
     1261 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1321 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1381 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1441 accgccgccg ggatcactct cggcatggac gagctgtaca agactagtga acagaaactg
     1501 attagcgaag aagatctgta aatcgatata actaagtaat atggtgcact ctcagtacaa
     1561 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     1621 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     1681 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggccgcc
     1741 cttcccaaca gttgcgcagc ctgaatggcg aatggcgctg atgtccggcg gtgcttttgc
     1801 cgttacgcac caccccgtca gtagctgaac aggagggaca gggggtgggc gaagaactcc
     1861 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     1921 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc
     1981 gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     2041 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     2101 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     2161 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     2221 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatcc
     2281 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     2341 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     2401 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     2461 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     2521 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     2581 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgg agttcattca
     2641 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     2701 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     2761 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     2821 ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     2881 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     2941 attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     3001 tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     3061 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc
     3121 tttagcagcc cttgcgccct gagtgcttgc ggcagcgtga agctgtgcct agaaatattt
     3181 tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc
     3241 tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc
     3301 tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta
     3361 gccttaaccg gcgcatgact tcaagactaa ctcctctaa
//
