LOCUS       sequence-266224-        6779 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 6779)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6779)
  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-281    C-Tag Venus173/155[1] L32 FRB EBPF2
FEATURES             Location/Qualifiers
     source          1..6779
                     /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"
     regulatory      1022..1031
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1033..1308
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1937..2511,2512..2514,2515..2523)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2104..2121)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2331..2350
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2524..2526,2527..2529,2530..3243)
                     /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(2569..2590)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3289..3421
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3430..4018
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3483..3503)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3901..3912)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4045..4062)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4051..4162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4258..4278)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4258..4276)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4297..4313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4297..4313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4310..4332)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4415..4434)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4480..4535
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4534..4553)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4914..5502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4994..5013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5673..6464,6465..6533))
                     /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     6296..6315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6534..6638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6706..6724
                     /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 accatggtgg ccatcctctg gcatgagatg tggcatgaag gcctggaaga ggcatctcgt
     1081 ttgtactttg gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct
     1141 atgatggaac ggggccccca gactctgaag gaaacatcct ttaatcaggc ctatggtcga
     1201 gatttaatgg aggcccaaga gtggtgcagg aagtacatga aatcagggaa tgtcaaggac
     1261 ctcacccaag cctgggacct ctattatcat gtgttccgac gaatctcaga cgtgggtggc
     1321 ggagggagcg aaggcggcgg atccggaggg cctggctctg gcggagaggg gagcgctgga
     1381 ggcgggtcag ccggtggtgg cagtgtgtca aagggagaag agttgtttac cggtgttgtt
     1441 cccatactgg tggaactgga tggtgacgtc aatggacaca agtttagcgt ttctggcgaa
     1501 ggagaaggcg atgctaccta tggcaaactg accctcaaac tgatctgtac gactggaaag
     1561 ttgcctgtac cttggccaac actggtcaca accttgggct atgggctgca atgctttgcc
     1621 cgttatcctg atcacatgaa gcaacatgac ttctttaagt ctgctatgcc agagggatat
     1681 gtgcaggaac ggactatctt cttcaaggac gacggcaact acaagacgag agctgaggtg
     1741 aaatttgagg gtgacacact ggtgaatcgc atagagctga aaggcatcga cttcaaagaa
     1801 gatgggaata ttctcgggca taaactcgag tacaactaca attcccacaa cgtgtacatc
     1861 actgccgata agcagaagaa tgggattaag gcgaacttca agatccgaca caacatcgag
     1921 taagaattca gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     1981 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     2041 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     2101 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     2161 tagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     2221 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     2281 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     2341 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     2401 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     2461 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     2521 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2581 gacggcgacg taaacggcca caagttcagc gtgaggggcg agggcgaggg cgatgccacc
     2641 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2701 accctcgtga ccaccctgag ccacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     2761 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2821 ttcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2881 ctggtgaacc gcatcgagct gaagggcgtc gacttcaagg aggacggcaa catcctgggg
     2941 cacaagctgg agtacaactt caacagccac aacatctata tcatggccgt caagcagaag
     3001 aacggcatca aggtgaactt caagatccgc cacaacgtgg aggacggcag cgtgcagctc
     3061 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacagc
     3121 cactacctga gcacccagtc cgtgctgagc aaagacccca acgagaagcg cgatcacatg
     3181 gtcctgctgg agttccgcac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3241 taaaccggtg ctcgaggtac cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag
     3301 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     3361 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     3421 ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3481 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3541 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3601 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3661 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3721 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3781 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     3841 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3901 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3961 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     4021 ggaattcgag ctcggtacga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4081 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4141 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4201 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcccagct tttgttccct
     4261 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4321 ttgttatccg ctaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     4381 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     4441 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     4501 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     4561 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     4621 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     4681 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     4741 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     4801 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     4861 cggccggccg ccggcgtcga cgcgcgcggc cgcggcgcgc cgttgctggc gtttttccat
     4921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6661 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6721 aggccctttc gtctacgcgc gcgccggcgg cgcgccgtcg acggccggcc gcggccgcg
//
