LOCUS       sequence-176355-        4701 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GF-ori-FP plasmid for GFxFP assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4701)
  AUTHORS   Toth E, Weinhardt N, Bencsura P, Huszar K, Kulcsar PI, Talas A, 
            Fodor E, Welker E
  TITLE     Cpf1 nucleases demonstrate robust activity to induce DNA 
            modification by exploiting homology directed repair pathways in 
            mammalian cells.
  JOURNAL   Biol Direct. 2016 Sep 14;11:46. doi: 10.1186/s13062-016-0147-0.
  PUBMED    27630115
REFERENCE   2  (bases 1 to 4701)
  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: "Biol 
            Direct."; date: "2016-09-14"; volume: "11:46. doi"; pages: " 
            10.1186/s13062-016-0147-0"
COMMENT     Sequence Label: pWN10042
FEATURES             Location/Qualifiers
     source          1..4701
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     308..330
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     390..409
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      437..446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(443..445,446..448,449..961)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(488..509)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     rep_origin      complement(1144..1732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1224..1243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2344..2365
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2494..2513)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2559..2614
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2613..2632)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2969..2988)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2970..2988)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2998..3015)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2998..3014)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3007..3029)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3156..3611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3243..3262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3453..3474
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3637..3741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3742..3810,3811..4602)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3960..3979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
       61 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
      121 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
      181 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
      241 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
      301 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gcctctgcta accatgttca
      361 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
      421 ttttggcaaa tctagagccg ccatggtgag caagggcgag gagctgttca ccggggtggt
      481 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      541 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      601 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      661 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      721 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      781 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      841 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      901 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
      961 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1021 cgtgctgctg cccgacaacc actgaggatc cgctagcctg caggtcgacg aattcgatga
     1081 tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1321 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1561 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1681 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1801 ggtcatgatc ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1861 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1921 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1981 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2041 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2101 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2161 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2221 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2281 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2341 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2401 gtacaagtaa ctcgagactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     2461 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     2521 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     2581 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     2641 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     2701 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     2761 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     2821 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     2881 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc
     2941 cctcgactta attaaggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     3001 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3061 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3121 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     3181 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3241 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3301 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3361 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3421 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3481 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3541 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3601 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3661 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3721 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3781 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3841 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3901 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3961 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4021 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4081 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4141 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4201 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4261 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4321 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4381 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4441 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4501 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4561 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4621 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4681 atcctttttg ataatctcat g
//
