LOCUS       sequence-249330-        6818 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 6818)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6818)
  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-219    C-Tag Venus173/155[1] L32 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6818
                     /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(1976..2550,2551..2553,2554..2562)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2143..2160)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2370..2389
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2563..2565,2566..2568,2569..3282)
                     /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(2608..2629)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3328..3460
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3469..4057
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3522..3542)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3940..3951)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4084..4101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4090..4201
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4297..4317)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4297..4315)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4336..4352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4336..4352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4349..4371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4454..4473)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4519..4574
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4573..4592)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4953..5541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5033..5052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5712..6503,6504..6572))
                     /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     6335..6354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6573..6677)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6745..6763
                     /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 aacatcgagt aagaattcag atccgcccct
     1981 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2041 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2101 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2161 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctggt agcttcttga agacaaacaa
     2221 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2281 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2341 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2401 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat
     2461 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     2521 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag
     2581 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2641 aagttcagcg tgaggggcga gggcgagggc gatgccacca acggcaagct gaccctgaag
     2701 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc
     2761 cacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2821 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcacc
     2881 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2941 aagggcgtcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc
     3001 aacagccaca acatctatat catggccgtc aagcagaaga acggcatcaa ggtgaacttc
     3061 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3121 acccccatcg gcgacggccc cgtgctgctg cccgacagcc actacctgag cacccagtcc
     3181 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttccgcacc
     3241 gccgccggga tcactctcgg catggacgag ctgtacaagt aaaccggtgc tcgaggtacc
     3301 gatgaatagc taaggtcgag gccgcaggta agtatcaagg ttacaagaca ggtttaagga
     3361 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     3421 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcgacaa tcaacctctg
     3481 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3541 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3601 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3661 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3721 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3781 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3841 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3901 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3961 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4021 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacgat
     4081 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4141 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4201 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4261 gggaggattg ggaagacaat agcccagctt ttgttccctt tagtgagggt taattgcgcg
     4321 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc taattcactc
     4381 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     4441 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     4501 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4561 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4621 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     4681 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4741 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4801 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     4861 tactcccagt catagctgtc cctcttctct tatggagatc ggccggccgc cggcgtcgac
     4921 gcgcgcggcc gcggcgcgcc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4981 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5041 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5101 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5161 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5221 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5281 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5341 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5401 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5461 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5521 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5581 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5641 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5701 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5761 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5821 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5881 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5941 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6001 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6061 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6121 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6181 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6241 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6301 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6361 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6421 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6481 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6541 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6601 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6661 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6721 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctacgcgcg
     6781 cgccggcggc gcgccgtcga cggccggccg cggccgcg
//
