LOCUS       sequence-249314-        6713 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 6713)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6713)
  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-277    C-Tag Venus173/155[1] L10 FRB EBPF2
FEATURES             Location/Qualifiers
     source          1..6713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     733..757
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1023..1032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1034..1309
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1872..2446,2447..2449,2450..2458)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2039..2056)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2266..2285
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2459..2461,2462..2464,2465..3178)
                     /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(2504..2525)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3224..3356
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3365..3953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3418..3438)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3836..3847)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3980..3997)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3986..4097
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4193..4213)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4193..4211)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4232..4248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4232..4248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4245..4267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4350..4369)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4415..4470
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4469..4488)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4849..5437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4929..4948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5608..6399,6400..6468))
                     /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     6231..6250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6469..6573)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6641..6659
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcctgcctcc aggcctcaat attggccatt agccatatta ttcattggtt atatagcata
       61 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
      121 tattggctca tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt
      781 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag
      841 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc
     1021 caccatggtg gccatcctct ggcatgagat gtggcatgaa ggcctggaag aggcatctcg
     1081 tttgtacttt ggggaaagga acgtgaaagg catgtttgag gtgctggagc ccttgcatgc
     1141 tatgatggaa cggggccccc agactctgaa ggaaacatcc tttaatcagg cctatggtcg
     1201 agatttaatg gaggcccaag agtggtgcag gaagtacatg aaatcaggga atgtcaagga
     1261 cctcacccaa gcctgggacc tctattatca tgtgttccga cgaatctcag ggagctctgc
     1321 cagcggtggc agcaccagtg tgtcaaaggg agaagagttg tttaccggtg ttgttcccat
     1381 actggtggaa ctggatggtg acgtcaatgg acacaagttt agcgtttctg gcgaaggaga
     1441 aggcgatgct acctatggca aactgaccct caaactgatc tgtacgactg gaaagttgcc
     1501 tgtaccttgg ccaacactgg tcacaacctt gggctatggg ctgcaatgct ttgcccgtta
     1561 tcctgatcac atgaagcaac atgacttctt taagtctgct atgccagagg gatatgtgca
     1621 ggaacggact atcttcttca aggacgacgg caactacaag acgagagctg aggtgaaatt
     1681 tgagggtgac acactggtga atcgcataga gctgaaaggc atcgacttca aagaagatgg
     1741 gaatattctc gggcataaac tcgagtacaa ctacaattcc cacaacgtgt acatcactgc
     1801 cgataagcag aagaatggga ttaaggcgaa cttcaagatc cgacacaaca tcgagtaaga
     1861 attcagatcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     1921 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     1981 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     2041 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggtagct
     2101 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     2161 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     2221 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     2281 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     2341 gggcctcggt acacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     2401 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     2461 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2521 cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc gagggcgatg ccaccaacgg
     2581 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2641 cgtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     2701 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2761 caaggacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2821 gaaccgcatc gagctgaagg gcgtcgactt caaggaggac ggcaacatcc tggggcacaa
     2881 gctggagtac aacttcaaca gccacaacat ctatatcatg gccgtcaagc agaagaacgg
     2941 catcaaggtg aacttcaaga tccgccacaa cgtggaggac ggcagcgtgc agctcgccga
     3001 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acagccacta
     3061 cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3121 gctggagttc cgcaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaac
     3181 cggtgctcga ggtaccgatg aatagctaag gtcgaggccg caggtaagta tcaaggttac
     3241 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
     3301 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     3361 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3421 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3481 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3541 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3601 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3661 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3721 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3781 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3841 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3901 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     3961 tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4021 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4081 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4141 gggcaggaca gcaaggggga ggattgggaa gacaatagcc cagcttttgt tccctttagt
     4201 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4261 atccgctaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     4321 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     4381 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     4441 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     4501 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     4561 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     4621 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     4681 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     4741 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatcggcc
     4801 ggccgccggc gtcgacgcgc gcggccgcgg cgcgccgttg ctggcgtttt tccataggct
     4861 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4921 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4981 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5041 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5101 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5161 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5221 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5281 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5521 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5581 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5641 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5701 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5761 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5821 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5881 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5941 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6001 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6061 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6121 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6181 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6241 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6301 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6361 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6421 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6601 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6661 ctttcgtcta cgcgcgcgcc ggcggcgcgc cgtcgacggc cggccgcggc cgc
//
