LOCUS       sequence-249358-        6344 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 6344)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6344)
  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-440    C-Tag Venus210[2] L20 FKBP mCherry
FEATURES             Location/Qualifiers
     source          1..6344
                     /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(1511..2085,2086..2088,2089..2097)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1678..1695)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1905..1924
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2098..2808
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2243..2261)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2511..2530
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2854..2986
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2995..3583
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3048..3068)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3466..3477)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3610..3627)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3616..3727
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3823..3843)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3823..3841)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3862..3878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3862..3878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3875..3897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3980..3999)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4045..4100
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4099..4118)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4479..5067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4559..4578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5238..6029,6030..6098))
                     /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     5861..5880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6099..6203)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6271..6289
                     /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 caccagtgat cccaatgaga agagggacca tatggtactg
     1441 ctcgaattcg tgactgcagc aggcattacc cttgggatgg atgaacttta taagtaagaa
     1501 ttcagatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     1561 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     1621 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     1681 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggtagctt
     1741 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     1801 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     1861 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     1921 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     1981 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2041 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     2101 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2161 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2221 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2281 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2341 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2401 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2461 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2521 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2581 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2641 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2701 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2761 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaac cggtgctcga
     2821 ggtaccgatg aatagctaag gtcgaggccg caggtaagta tcaaggttac aagacaggtt
     2881 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     2941 gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt cgacaatcaa
     3001 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3061 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3121 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3181 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3241 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3301 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3361 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     3421 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3481 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3541 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg
     3601 tacgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3661 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3721 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3781 gcaaggggga ggattgggaa gacaatagcc cagcttttgt tccctttagt gagggttaat
     3841 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctaat
     3901 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     3961 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     4021 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     4081 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     4141 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     4201 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     4261 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     4321 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     4381 cctgactact cccagtcata gctgtccctc ttctcttatg gagatcggcc ggccgccggc
     4441 gtcgacgcgc gcggccgcgg cgcgccgttg ctggcgtttt tccataggct ccgcccccct
     4501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6241 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcta
     6301 cgcgcgcgcc ggcggcgcgc cgtcgacggc cggccgcggc cgcg
//
