LOCUS       sequence-194133-        3045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCYFP. Expresses CYFP control fusion in E. coli for TriFC assay 
            (kanR)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3045)
  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 3045)
  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 220
FEATURES             Location/Qualifiers
     source          1..3045
                     /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             877..1128
                     /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     1065..1086
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1135..1164
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1202..1221)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1321..1343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1661..2455)
                     /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(1772..1791)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2382..2401
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      join(2851..3045,1..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 gcttgaaaca gaaagttatg aatcatgaca agcagaagaa cggcatcaag
      901 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
      961 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1021 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1081 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagac tagtgaacag
     1141 aaactgatta gcgaagaaga tctgtaaatc gatataacta agtaatatgg tgcactctca
     1201 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     1261 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     1321 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     1381 gccgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctgatgt ccggcggtgc
     1441 ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga gggacagggg gtgggcgaag
     1501 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
     1561 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
     1621 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     1681 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     1741 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     1801 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     1861 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     1921 gcatccgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     1981 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     2041 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     2101 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     2161 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     2221 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttggagtt
     2281 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     2341 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     2401 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     2461 acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg
     2521 gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag
     2581 ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
     2641 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
     2701 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc
     2761 gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgaagct gtgcctagaa
     2821 atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc
     2881 ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac
     2941 caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc
     3001 agtttagcct taaccggcgc atgacttcaa gactaactcc tctaa
//
