LOCUS       sequence-249371-        6275 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 6275)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6275)
  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-139    N-Tag GFP[1] L20 FRB mCherry
FEATURES             Location/Qualifiers
     source          1..6275
                     /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             1151..1426
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1443..2017,2018..2020,2021..2029)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1610..1627)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1837..1856
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2030..2740
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2175..2193)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2443..2462
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2786..2918
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2927..3515
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2980..3000)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3398..3409)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3542..3559)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3548..3659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3755..3775)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3755..3773)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3794..3810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3794..3810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3807..3829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3912..3931)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3977..4032
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4031..4050)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4411..4999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4491..4510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5170..5961,5962..6030))
                     /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     5793..5812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6031..6135)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6203..6221
                     /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 caccatggat ctgcctgatg accactacct gagcacacag accatcctct ccaaggacct
     1081 taacgggagt cctggcggtg gatccggagg ctctgggagc tctgccagcg gtggcagcac
     1141 ctccgtggcc atcctctggc atgagatgtg gcatgaaggc ctggaagagg catctcgttt
     1201 gtactttggg gaaaggaacg tgaaaggcat gtttgaggtg ctggagccct tgcatgctat
     1261 gatggaacgg ggcccccaga ctctgaagga aacatccttt aatcaggcct atggtcgaga
     1321 tttaatggag gcccaagagt ggtgcaggaa gtacatgaaa tcagggaatg tcaaggacct
     1381 cacccaagcc tgggacctct attatcatgt gttccgacga atctcataag aattcagatc
     1441 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     1501 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     1561 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     1621 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggtagc ttcttgaaga
     1681 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     1741 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     1801 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     1861 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     1921 tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     1981 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa
     2041 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     2101 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2161 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     2221 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     2281 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2341 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     2401 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     2461 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     2521 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     2581 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     2641 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     2701 ccgccactcc accggcggca tggacgagct gtacaagtaa accggtgctc gaggtaccga
     2761 tgaatagcta aggtcgaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga
     2821 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     2881 tggtcttact gacatccact ttgcctttct ctccacaggt gtcgacaatc aacctctgga
     2941 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3001 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3061 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3121 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3181 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3241 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3301 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3361 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3421 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3481 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc ggtacgatca
     3541 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3601 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3661 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3721 gaggattggg aagacaatag cccagctttt gttcccttta gtgagggtta attgcgcgct
     3781 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgcta attcactcct
     3841 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     3901 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     3961 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     4021 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     4081 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     4141 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     4201 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4261 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4321 ctcccagtca tagctgtccc tcttctctta tggagatcgg ccggccgccg gcgtcgacgc
     4381 gcgcggccgc ggcgcgccgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4441 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4501 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4561 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4621 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4681 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4741 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4801 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4861 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4921 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4981 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5041 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5101 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5161 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5221 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5281 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5341 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5401 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5461 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5521 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5581 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5641 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5701 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5761 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5821 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5881 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5941 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6001 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6061 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6121 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6181 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tacgcgcgcg
     6241 ccggcggcgc gccgtcgacg gccggccgcg gccgc
//
