LOCUS       sequence-249307-        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-462    N-Tag UnaG[1] L32 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6551
                     /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             1373..1696
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1710..2284,2285..2287,2288..2296)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1877..1894)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2104..2123
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2297..2299,2300..2302,2303..3016)
                     /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(2342..2363)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3062..3194
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3203..3791
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3256..3276)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3674..3685)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3818..3835)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3824..3935
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4031..4051)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4031..4049)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4070..4086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4070..4086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4083..4105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4188..4207)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4253..4308
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4307..4326)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4687..5275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4767..4786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5446..6237,6238..6306))
                     /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     6069..6088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6307..6411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6479..6497
                     /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 aggaaagacg tgggtggcgg agggagcgaa ggcggcggat ccggagggcc
     1321 tggctctggc ggagagggga gcgctggagg cgggtcagcc ggtggtggga gtggagtgca
     1381 ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt
     1441 ggtgcactac accgggatgc ttgaagatgg aaagaaattt gattcctccc gggacagaaa
     1501 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
     1561 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
     1621 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
     1681 tctaaaactg gaataagaat tcagatccgc ccctctccct cccccccccc taacgttact
     1741 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     1801 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     1861 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     1921 gcagttcctc tggtagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     1981 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     2041 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     2101 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     2161 tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta
     2221 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     2281 taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2341 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga
     2401 gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2461 cgtgccctgg cccaccctcg tgaccaccct gagccacggc gtgcagtgct tcgcccgcta
     2521 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2581 ggagcgcacc atcttcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt
     2641 cgagggcgac accctggtga accgcatcga gctgaagggc gtcgacttca aggaggacgg
     2701 caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacatct atatcatggc
     2761 cgtcaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg
     2821 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2881 gctgcccgac agccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa
     2941 gcgcgatcac atggtcctgc tggagttccg caccgccgcc gggatcactc tcggcatgga
     3001 cgagctgtac aagtaaaccg gtgctcgagg taccgatgaa tagctaaggt cgaggccgca
     3061 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
     3121 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
     3181 ctttctctcc acaggtgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3241 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3301 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3361 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3421 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3481 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3541 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3601 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3661 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3721 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3781 cgcctccccg cctggaattc gagctcggta cgatcagcct cgactgtgcc ttctagttgc
     3841 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3901 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3961 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagccca
     4021 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4081 ttcctgtgtg aaattgttat ccgctaattc actcctcagg tgcaggctgc ctatcagaag
     4141 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     4201 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     4261 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     4321 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     4381 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     4441 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     4501 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     4561 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     4621 ctcttatgga gatcggccgg ccgccggcgt cgacgcgcgc ggccgcggcg cgccgttgct
     4681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5101 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5221 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5281 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5341 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5401 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5461 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5521 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5581 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5641 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5701 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5761 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5821 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5881 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5941 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6001 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6061 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6121 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6181 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6241 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6301 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6361 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6421 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6481 taggcgtatc acgaggccct ttcgtctacg cgcgcgccgg cggcgcgccg tcgacggccg
     6541 gccgcggccg c
//
