LOCUS       sequence-266222-        6515 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 6515)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6515)
  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-460    N-Tag UnaG[1] L20 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6515
                     /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"
     CDS             1337..1660
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1674..2248,2249..2251,2252..2260)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1841..1858)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2068..2087
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2261..2263,2264..2266,2267..2980)
                     /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(2306..2327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3026..3158
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3167..3755
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3220..3240)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3638..3649)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3782..3799)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3788..3899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3995..4015)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3995..4013)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4034..4050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4034..4050)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4047..4069)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4152..4171)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4217..4272
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4271..4290)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4651..5239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4731..4750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5410..6201,6202..6270))
                     /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     6033..6052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6271..6375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6443..6461
                     /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 caccatggtc gaaaagttcg tgggcacatg gaagattgcc gatagccaca acttcggcga
     1081 atacctcaaa gctatagggg caccaaaaga gcttagcgat ggtggagatg cgacaactcc
     1141 cacactgtac attagccaga aagacggaga caagatgact gtcaagatcg agaatgggcc
     1201 tcccaccttt ctggacacac aggttaagtt caaactgggt gaggagtttg acgagtttcc
     1261 cagtgatcgc aggaaaggga gtcctggcgg tggatccgga ggctctggga gctctgccag
     1321 cggtggcagc acctccggag tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt
     1381 ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa
     1441 atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt
     1501 gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac
     1561 tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc
     1621 cactctcgtc ttcgatgtgg agcttctaaa actggaataa gaattcagat ccgcccctct
     1681 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     1741 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     1801 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     1861 ggtctgttga atgtcgtgaa ggaagcagtt cctctggtag cttcttgaag acaaacaacg
     1921 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     1981 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     2041 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     2101 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc
     2161 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     2221 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     2281 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2341 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     2401 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
     2461 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2521 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
     2581 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2641 gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     2701 cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa
     2761 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2821 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
     2881 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc
     2941 cgccgggatc actctcggca tggacgagct gtacaagtaa accggtgctc gaggtaccga
     3001 tgaatagcta aggtagaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga
     3061 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     3121 tggtcttact gacatccact ttgcctttct ctccacaggt gtcgacaatc aacctctgga
     3181 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3241 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3301 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3361 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3421 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3481 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3541 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3601 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3661 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3721 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc ggtacgatca
     3781 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3841 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3901 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3961 gaggattggg aagacaatag cccagctttt gttcccttta gtgagggtta attgcgcgct
     4021 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgcta attcactcct
     4081 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     4141 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     4201 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     4261 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     4321 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     4381 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     4441 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4501 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4561 ctcccagtca tagctgtccc tcttctctta tggagatcgg ccggccgccg gcgtcgacgc
     4621 gcgcggccgc ggcgcgccgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5161 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5341 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5401 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5461 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5521 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5581 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5641 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5701 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5761 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5821 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5881 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5941 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6001 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6061 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6121 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6181 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6241 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6301 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6361 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6421 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tacgcgcgcg
     6481 ccggcggcgc gccgtcgacg gccggccgcg gccgc
//
