LOCUS       sequence-249352-        6893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PPI (BiFC/TriFC) assay vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6893)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6893)
  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"
COMMENT     Sequence Label: pRDAX-434    C-Tag Venus210[1] L20 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1018..1027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1024..1026,1027..1347)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     misc_feature    join(2051..2625,2626..2628,2629..2637)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2218..2235)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2445..2464
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2638..2640,2641..2643,2644..3357)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(2683..2704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3403..3535
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3544..4132
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3597..3617)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4015..4026)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4159..4176)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4165..4276
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4372..4392)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4372..4390)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4411..4427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4411..4427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4424..4446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4529..4548)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4594..4649
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4648..4667)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5028..5616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5108..5127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5787..6578,6579..6647))
                     /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     6410..6429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6648..6752)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6820..6838
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctgcctcca ggcctcaata ttggccatta gccatattat tcattggtta tatagcataa
       61 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      121 attggctcat gtccaatatg accgccatgt tggcattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggcc
     1021 accatgggag tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc
     1081 ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa atttgattcc
     1141 tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc
     1201 tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca
     1261 gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc
     1321 ttcgatgtgg agcttctaaa actggaaggg agtcctggcg gtggatccgg aggctctggg
     1381 agctctgcca gcggtggcag caccagtgtg tcaaagggag aagagttgtt taccggtgtt
     1441 gttcccatac tggtggaact ggatggtgac gtcaatggac acaagtttag cgtttctggc
     1501 gaaggagaag gcgatgctac ctatggcaaa ctgaccctca aactgatctg tacgactgga
     1561 aagttgcctg taccttggcc aacactggtc acaaccttgg gctatgggct gcaatgcttt
     1621 gcccgttatc ctgatcacat gaagcaacat gacttcttta agtctgctat gccagaggga
     1681 tatgtgcagg aacggactat cttcttcaag gacgacggca actacaagac gagagctgag
     1741 gtgaaatttg agggtgacac actggtgaat cgcatagagc tgaaaggcat cgacttcaaa
     1801 gaagatggga atattctcgg gcataaactc gagtacaact acaattccca caacgtgtac
     1861 atcactgccg ataagcagaa gaatgggatt aaggcgaact tcaagatccg acacaacatc
     1921 gaggatggtg gggtccagct tgccgaccat tatcagcaga acacacccat tggggatgga
     1981 ccagtcctgt tgccggataa ccactacctc agttaccaga gcaaactgtc caaataagaa
     2041 ttcagatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     2101 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     2161 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     2221 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggtagctt
     2281 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     2341 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     2401 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2461 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2521 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2581 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     2641 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2701 gacgtaaacg gccacaagtt cagcgtgagg ggcgagggcg agggcgatgc caccaacggc
     2761 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2821 gtgaccaccc tgagccacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     2881 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2941 aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3001 aaccgcatcg agctgaaggg cgtcgacttc aaggaggacg gcaacatcct ggggcacaag
     3061 ctggagtaca acttcaacag ccacaacatc tatatcatgg ccgtcaagca gaagaacggc
     3121 atcaaggtga acttcaagat ccgccacaac gtggaggacg gcagcgtgca gctcgccgac
     3181 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga cagccactac
     3241 ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3301 ctggagttcc gcaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaacc
     3361 ggtgctcgag gtaccgatga atagctaagg tcgaggccgc aggtaagtat caaggttaca
     3421 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
     3481 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
     3541 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3601 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3661 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3721 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3781 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3841 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3901 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     3961 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4021 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4081 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     4141 cgagctcggt acgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4201 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4261 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4321 ggcaggacag caagggggag gattgggaag acaatagccc agcttttgtt ccctttagtg
     4381 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4441 tccgctaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     4501 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     4561 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     4621 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     4681 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     4741 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     4801 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     4861 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     4921 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatcggccg
     4981 gccgccggcg tcgacgcgcg cggccgcggc gcgccgttgc tggcgttttt ccataggctc
     5041 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5101 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5161 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5221 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5281 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5341 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5401 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5461 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5521 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5581 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5641 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5701 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5761 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5821 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5881 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5941 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6001 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6061 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6121 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6181 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6241 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6301 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6361 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6421 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6481 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6541 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6601 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6661 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6721 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6781 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6841 tttcgtctac gcgcgcgccg gcggcgcgcc gtcgacggcc ggccgcggcc gcg
//
