LOCUS       sequence-249280-        6641 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 6641)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6641)
  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-113    C-Tag miRFP670[1] L20 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6641
                     /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(1799..2373,2374..2376,2377..2385)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1966..1983)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2193..2212
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2386..2388,2389..2391,2392..3105)
                     /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(2431..2452)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3151..3283
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3292..3880
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3345..3365)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3763..3774)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3907..3924)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3913..4024
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4120..4140)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4120..4138)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4159..4175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4159..4175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4172..4194)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4277..4296)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4342..4397
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4396..4415)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4776..5364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4856..4875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5535..6326,6327..6395))
                     /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     6158..6177
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6396..6500)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6568..6586
                     /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 actggaaggg agtcctggcg gtggatccgg aggctctggg
     1381 agctctgcca gcggtggcag caccagtgtg gctgggcatg ctagcggttc tcctgccttt
     1441 ggcaccgcta gtcactccaa ctgcgagcac gaggagatcc atcttgctgg gagtatccag
     1501 cctcacggag cccttctggt cgttagcgaa cacgatcacc gtgtgattca ggcaagcgcg
     1561 aatgcagccg agttcctcaa cctgggcagt gtgctgggag ttcccttggc cgaaatcgac
     1621 ggagatctgc tgatcaagat tctgccccat ctcgatccga cagctgaagg catgccagta
     1681 gcagttaggt gtcgcattgg gaatccgtcc accgagtatt gcggtctgat gcataggcca
     1741 ccagaaggag ggctgattat cgagctggaa agagcgggac cctaagaatt cagatccgcc
     1801 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     1861 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     1921 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     1981 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggtagcttct tgaagacaaa
     2041 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     2101 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     2161 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     2221 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca
     2281 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     2341 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc
     2401 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2461 cacaagttca gcgtgagggg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     2521 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2581 agccacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2641 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2701 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2761 ctgaagggcg tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2821 ttcaacagcc acaacatcta tatcatggcc gtcaagcaga agaacggcat caaggtgaac
     2881 ttcaagatcc gccacaacgt ggaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2941 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca gccactacct gagcacccag
     3001 tccgtgctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttccgc
     3061 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaaccgg tgctcgaggt
     3121 accgatgaat agctaaggtc gaggccgcag gtaagtatca aggttacaag acaggtttaa
     3181 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     3241 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcga caatcaacct
     3301 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3361 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3421 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3481 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3541 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3601 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3661 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     3721 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3781 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3841 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     3901 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3961 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4021 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4081 agggggagga ttgggaagac aatagcccag cttttgttcc ctttagtgag ggttaattgc
     4141 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctaattca
     4201 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     4261 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     4321 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     4381 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     4441 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     4501 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     4561 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     4621 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     4681 gactactccc agtcatagct gtccctcttc tcttatggag atcggccggc cgccggcgtc
     4741 gacgcgcgcg gccgcggcgc gccgttgctg gcgtttttcc ataggctccg cccccctgac
     4801 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4861 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4921 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4981 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5041 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5101 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5161 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5221 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5281 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5341 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5401 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5461 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5521 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5581 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5641 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5701 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5761 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5821 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5881 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5941 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6001 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6061 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6121 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6181 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6241 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6301 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6361 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6421 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6481 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6541 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctacgc
     6601 gcgcgccggc ggcgcgccgt cgacggccgg ccgcggccgc g
//
