LOCUS       sequence-249340-        6854 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 6854)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6854)
  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-446    C-Tag Venus210[1] L20 FRB EBPF2
FEATURES             Location/Qualifiers
     source          1..6854
                     /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(2012..2586,2587..2589,2590..2598)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2179..2196)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2406..2425
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2599..2601,2602..2604,2605..3318)
                     /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(2644..2665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3364..3496
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3505..4093
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3558..3578)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3976..3987)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4120..4137)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4126..4237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4333..4353)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4333..4351)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4372..4388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4372..4388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4385..4407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4490..4509)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4555..4610
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4609..4628)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4989..5577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5069..5088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5748..6539,6540..6608))
                     /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     6371..6390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6609..6713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6781..6799
                     /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 cgaggatggt ggggtccagc ttgccgacca ttatcagcag
     1921 aacacaccca ttggggatgg accagtcctg ttgccggata accactacct cagttaccag
     1981 agcaaactgt ccaaataaga attcagatcc gcccctctcc ctcccccccc cctaacgtta
     2041 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     2101 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     2161 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2221 aagcagttcc tctggtagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     2281 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     2341 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     2401 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     2461 attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt tagtcgaggt
     2521 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     2581 gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2641 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc
     2701 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2761 cccgtgccct ggcccaccct cgtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc
     2821 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2881 caggagcgca ccatcttctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     2941 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcgtcgactt caaggaggac
     3001 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacat ctatatcatg
     3061 gccgtcaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa cgtggaggac
     3121 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3181 ctgctgcccg acagccacta cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag
     3241 aagcgcgatc acatggtcct gctggagttc cgcaccgccg ccgggatcac tctcggcatg
     3301 gacgagctgt acaagtaaac cggtgctcga ggtaccgatg aatagctaag gtcgaggccg
     3361 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
     3421 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     3481 gcctttctct ccacaggtgt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     3541 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3601 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3661 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3721 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3781 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3841 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     3901 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3961 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4021 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4081 gccgcctccc cgcctggaat tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt
     4141 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4201 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4261 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagcc
     4321 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     4381 gtttcctgtg tgaaattgtt atccgctaat tcactcctca ggtgcaggct gcctatcaga
     4441 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     4501 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     4561 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     4621 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     4681 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     4741 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     4801 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     4861 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     4921 ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc gcggccgcgg cgcgccgttg
     4981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5401 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5641 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5701 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5761 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5821 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5881 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5941 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6001 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6061 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6181 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6481 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6601 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6721 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6781 aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc ggcggcgcgc cgtcgacggc
     6841 cggccgcggc cgcg
//
