LOCUS       sequence-249306-        6551 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 6551)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6551)
  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-461    C-Tag UnaG[1] L32 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6551
                     /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(1709..2283,2284..2286,2287..2295)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1876..1893)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2103..2122
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2296..2298,2299..2301,2302..3015)
                     /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(2341..2362)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3061..3193
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3202..3790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3255..3275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3673..3684)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3817..3834)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3823..3934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4030..4050)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4030..4048)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4069..4085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4069..4085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4082..4104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4187..4206)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4252..4307
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4306..4325)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4686..5274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4766..4785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5445..6236,6237..6305))
                     /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     6068..6087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6306..6410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6478..6496
                     /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 cggtggtggg
     1441 agtgtcgaaa agttcgtggg cacatggaag attgccgata gccacaactt cggcgaatac
     1501 ctcaaagcta taggggcacc aaaagagctt agcgatggtg gagatgcgac aactcccaca
     1561 ctgtacatta gccagaaaga cggagacaag atgactgtca agatcgagaa tgggcctccc
     1621 acctttctgg acacacaggt taagttcaaa ctgggtgagg agtttgacga gtttcccagt
     1681 gatcgcagga aataagaatt cagatccgcc cctctccctc ccccccccct aacgttactg
     1741 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     1801 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     1861 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     1921 cagttcctct ggtagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     1981 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     2041 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     2101 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     2161 gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa
     2221 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     2281 aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2341 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgagggg cgagggcgag
     2401 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2461 gtgccctggc ccaccctcgt gaccaccctg agccacggcg tgcagtgctt cgcccgctac
     2521 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2581 gagcgcacca tcttcttcaa ggacgacggc acctacaaga cccgcgccga ggtgaagttc
     2641 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggcg tcgacttcaa ggaggacggc
     2701 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacatcta tatcatggcc
     2761 gtcaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacgt ggaggacggc
     2821 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2881 ctgcccgaca gccactacct gagcacccag tccgtgctga gcaaagaccc caacgagaag
     2941 cgcgatcaca tggtcctgct ggagttccgc accgccgccg ggatcactct cggcatggac
     3001 gagctgtaca agtaaaccgg tgctcgaggt accgatgaat agctaaggtc gaggccgcag
     3061 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
     3121 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     3181 tttctctcca caggtgtcga caatcaacct ctggattaca aaatttgtga aagattgact
     3241 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3301 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3361 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3421 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3481 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3541 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     3601 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3661 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3721 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3781 gcctccccgc ctggaattcg agctcggtac gatcagcctc gactgtgcct tctagttgcc
     3841 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3901 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3961 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcccag
     4021 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     4081 tcctgtgtga aattgttatc cgctaattca ctcctcaggt gcaggctgcc tatcagaagg
     4141 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     4201 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     4261 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4321 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4381 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     4441 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     4501 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     4561 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     4621 tcttatggag atcggccggc cgccggcgtc gacgcgcgcg gccgcggcgc gccgttgctg
     4681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4861 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5101 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5281 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5761 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6061 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6421 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6481 aggcgtatca cgaggccctt tcgtctacgc gcgcgccggc ggcgcgccgt cgacggccgg
     6541 ccgcggccgc g
//
