LOCUS       sequence-249342-        6890 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 6890)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6890)
  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-448    C-Tag Venus210[1] L32 FRB EBPF2
FEATURES             Location/Qualifiers
     source          1..6890
                     /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(2048..2622,2623..2625,2626..2634)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2215..2232)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2442..2461
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2635..2637,2638..2640,2641..3354)
                     /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(2680..2701)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3400..3532
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3541..4129
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3594..3614)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4012..4023)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4156..4173)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4162..4273
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4369..4389)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4369..4387)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4408..4424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4408..4424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4421..4443)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4526..4545)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4591..4646
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4645..4664)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5025..5613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5105..5124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5784..6575,6576..6644))
                     /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     6407..6426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6645..6749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6817..6835
                     /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 gatggtgggg tccagcttgc cgaccattat cagcagaaca cacccattgg ggatggacca
     1981 gtcctgttgc cggataacca ctacctcagt taccagagca aactgtccaa ataagaattc
     2041 agatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     2101 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     2161 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     2221 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt
     2281 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     2341 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     2401 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     2461 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     2521 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     2581 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     2641 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2701 gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac caacggcaag
     2761 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2821 accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     2881 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2941 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3001 cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg gcacaagctg
     3061 gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa gaacggcatc
     3121 aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac
     3181 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag ccactacctg
     3241 agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3301 gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaccggt
     3361 gctcgaggta ccgatgaata gctaaggtcg aggccgcagg taagtatcaa ggttacaaga
     3421 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     3481 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtcgac
     3541 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3601 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3661 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3721 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3781 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3841 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3901 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3961 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4021 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4081 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
     4141 gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4201 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4261 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4321 aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc tttagtgagg
     4381 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4441 gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     4501 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     4561 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     4621 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     4681 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     4741 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     4801 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     4861 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     4921 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tcggccggcc
     4981 gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca taggctccgc
     5041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5461 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5761 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5821 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5881 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5941 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6001 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6061 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6121 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6181 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6241 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6301 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6361 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6421 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6481 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6541 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6601 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6661 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6721 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6781 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6841 cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc cgcggccgcg
//
