LOCUS       sequence-123456-        1121 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  TOPO construct expressing CFP for generation of GMAP-compatible 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1121)
  AUTHORS   Akama-Garren EH, Joshi NS, Tammela T, Chang GP, Wagner BL, Lee DY, 
            Rideout Iii WM, Papagiannakopoulos T, Xue W, Jacks T
  TITLE     A Modular Assembly Platform for Rapid Generation of DNA Constructs.
  JOURNAL   Sci Rep. 2016 Feb 18;6:16836. doi: 10.1038/srep16836.
  PUBMED    26887506
REFERENCE   2  (bases 1 to 1121)
  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: "Sci Rep."; 
            date: "2016-02-18"; volume: "6:16836. doi"; pages: " 
            10.1038/srep16836"
COMMENT     Sequence Label: TOPO CFP
FEATURES             Location/Qualifiers
     source          1..1121
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     210..227
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    344..365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(380..385,386..403,404..410)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    418..434
                     /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     423..445
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     442..458
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     442..458
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        476..494
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     476..493
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(569..571,572..574,575..1087)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(614..635)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(875..894)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
       61 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      121 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      181 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      241 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      301 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      361 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      421 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctattta
      481 ggtgacacta tagaatactc aagctatgca tcaagcttgg taccgagctc ggatcctaac
      541 tcgaacgcta gctgtgcgat cgtttaccat ggtgagcaag ggcgaggagc tgttcaccgg
      601 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      661 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
      721 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg
      781 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      841 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
      901 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
      961 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt
     1021 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1081 catcgaggac ggcagcgtgc agctcgccga ccactaccag c
//
