LOCUS       sequence-271312-        6743 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 6743)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6743)
  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-279    C-Tag Venus173/155[1] L20 FRB EBPF2
FEATURES             Location/Qualifiers
     source          1..6743
                     /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(1901..2475,2476..2478,2479..2487)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2068..2085)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2295..2314
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2488..2490,2491..2493,2494..3207)
                     /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(2533..2554)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3253..3385
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3394..3982
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3447..3467)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3865..3876)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4009..4026)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4015..4126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4222..4242)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4222..4240)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4261..4277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4261..4277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4274..4296)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4379..4398)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4444..4499
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4498..4517)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4878..5466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4958..4977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5637..6428,6429..6497))
                     /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     6260..6279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6498..6602)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6670..6688
                     /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 gaatctcagg gagtcctggc
     1321 ggtggatccg gaggctctgg gagctctgcc agcggtggca gcaccagtgt gtcaaaggga
     1381 gaagagttgt ttaccggtgt tgttcccata ctggtggaac tggatggtga cgtcaatgga
     1441 cacaagttta gcgtttctgg cgaaggagaa ggcgatgcta cctatggcaa actgaccctc
     1501 aaactgatct gtacgactgg aaagttgcct gtaccttggc caacactggt cacaaccttg
     1561 ggctatgggc tgcaatgctt tgcccgttat cctgatcaca tgaagcaaca tgacttcttt
     1621 aagtctgcta tgccagaggg atatgtgcag gaacggacta tcttcttcaa ggacgacggc
     1681 aactacaaga cgagagctga ggtgaaattt gagggtgaca cactggtgaa tcgcatagag
     1741 ctgaaaggca tcgacttcaa agaagatggg aatattctcg ggcataaact cgagtacaac
     1801 tacaattccc acaacgtgta catcactgcc gataagcaga agaatgggat taaggcgaac
     1861 ttcaagatcc gacacaacat cgagtaagaa ttcagatccg cccctctccc tccccccccc
     1921 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     1981 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     2041 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     2101 tcgtgaagga agcagttcct ctggtagctt cttgaagaca aacaacgtct gtagcgaccc
     2161 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     2221 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     2281 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     2341 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
     2401 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     2461 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagct gttcaccggg
     2521 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgagg
     2581 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc
     2641 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgagccacgg cgtgcagtgc
     2701 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2761 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcacctacaa gacccgcgcc
     2821 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg cgtcgacttc
     2881 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaacatc
     2941 tatatcatgg ccgtcaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3001 gtggaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3061 ggccccgtgc tgctgcccga cagccactac ctgagcaccc agtccgtgct gagcaaagac
     3121 cccaacgaga agcgcgatca catggtcctg ctggagttcc gcaccgccgc cgggatcact
     3181 ctcggcatgg acgagctgta caagtaaacc ggtgctcgag gtaccgatga atagctaagg
     3241 tcgaggccgc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
     3301 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
     3361 atccactttg cctttctctc cacaggtgtc gacaatcaac ctctggatta caaaatttgt
     3421 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3481 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3541 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3601 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3661 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3721 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3781 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     3841 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3901 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3961 tccctttggg ccgcctcccc gcctggaatt cgagctcggt acgatcagcc tcgactgtgc
     4021 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4081 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4141 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4201 acaatagccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     4261 gtcatagctg tttcctgtgt gaaattgtta tccgctaatt cactcctcag gtgcaggctg
     4321 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     4381 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4441 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4501 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4561 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     4621 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     4681 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     4741 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     4801 ctgtccctct tctcttatgg agatcggccg gccgccggcg tcgacgcgcg cggccgcggc
     4861 gcgccgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4921 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4981 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5041 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5101 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5221 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5281 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5341 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5461 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5521 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5761 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6601 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6661 acctataaaa ataggcgtat cacgaggccc tttcgtctac gcgcgcgccg gcggcgcgcc
     6721 gtcgacggcc ggccgcggcc gcg
//
