LOCUS       sequence-249354-        6929 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 6929)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6929)
  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-436    C-Tag Venus210[1] L32 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6929
                     /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(2087..2661,2662..2664,2665..2673)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2254..2271)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2481..2500
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2674..2676,2677..2679,2680..3393)
                     /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(2719..2740)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3439..3571
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3580..4168
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3633..3653)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4051..4062)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4195..4212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4201..4312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4408..4428)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4408..4426)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4447..4463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4447..4463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4460..4482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4565..4584)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4630..4685
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4684..4703)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5064..5652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5144..5163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5823..6614,6615..6683))
                     /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     6446..6465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6684..6788)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6856..6874
                     /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 actggaagac gtgggtggcg gagggagcga aggcggcgga
     1381 tccggagggc ctggctctgg cggagagggg agcgctggag gcgggtcagc cggtggtggc
     1441 agtgtgtcaa agggagaaga gttgtttacc ggtgttgttc ccatactggt ggaactggat
     1501 ggtgacgtca atggacacaa gtttagcgtt tctggcgaag gagaaggcga tgctacctat
     1561 ggcaaactga ccctcaaact gatctgtacg actggaaagt tgcctgtacc ttggccaaca
     1621 ctggtcacaa ccttgggcta tgggctgcaa tgctttgccc gttatcctga tcacatgaag
     1681 caacatgact tctttaagtc tgctatgcca gagggatatg tgcaggaacg gactatcttc
     1741 ttcaaggacg acggcaacta caagacgaga gctgaggtga aatttgaggg tgacacactg
     1801 gtgaatcgca tagagctgaa aggcatcgac ttcaaagaag atgggaatat tctcgggcat
     1861 aaactcgagt acaactacaa ttcccacaac gtgtacatca ctgccgataa gcagaagaat
     1921 gggattaagg cgaacttcaa gatccgacac aacatcgagg atggtggggt ccagcttgcc
     1981 gaccattatc agcagaacac acccattggg gatggaccag tcctgttgcc ggataaccac
     2041 tacctcagtt accagagcaa actgtccaaa taagaattca gatccgcccc tctccctccc
     2101 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     2161 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     2221 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     2281 tgaatgtcgt gaaggaagca gttcctctgg tagcttcttg aagacaaaca acgtctgtag
     2341 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     2401 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     2461 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     2521 cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca tgctttacat
     2581 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     2641 tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga ggagctgttc
     2701 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2761 gtgaggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc
     2821 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgag ccacggcgtg
     2881 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2941 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcac ctacaagacc
     3001 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcgtc
     3061 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac
     3121 aacatctata tcatggccgt caagcagaag aacggcatca aggtgaactt caagatccgc
     3181 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3241 ggcgacggcc ccgtgctgct gcccgacagc cactacctga gcacccagtc cgtgctgagc
     3301 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttccgcac cgccgccggg
     3361 atcactctcg gcatggacga gctgtacaag taaaccggtg ctcgaggtac cgatgaatag
     3421 ctaaggtcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
     3481 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     3541 actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa
     3601 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3661 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3721 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3781 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3841 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3901 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3961 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4021 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4081 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4141 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga
     4201 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4261 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4321 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4381 gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     4441 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact cctcaggtgc
     4501 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     4561 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     4621 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     4681 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     4741 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     4801 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     4861 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     4921 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     4981 tcatagctgt ccctcttctc ttatggagat cggccggccg ccggcgtcga cgcgcgcggc
     5041 cgcggcgcgc cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6781 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6841 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctacgcgc gcgccggcgg
     6901 cgcgccgtcg acggccggcc gcggccgcg
//
