LOCUS       sequence-249369-        6245 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 6245)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6245)
  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-110    N-Tag GFP[1] L10 FRB mCherry
FEATURES             Location/Qualifiers
     source          1..6245
                     /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             1121..1396
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1413..1987,1988..1990,1991..1999)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1580..1597)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1807..1826
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2000..2710
                     /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(2145..2163)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2413..2432
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2756..2888
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2897..3485
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2950..2970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3368..3379)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3512..3529)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3518..3629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3725..3745)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3725..3743)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3764..3780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3764..3780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3777..3799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3882..3901)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3947..4002
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4001..4020)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4381..4969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4461..4480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5140..5931,5932..6000))
                     /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     5763..5782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6001..6105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6173..6191
                     /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 taacgggagc tctgccagcg gtggcagcac ctccgtggcc atcctctggc atgagatgtg
     1141 gcatgaaggc ctggaagagg catctcgttt gtactttggg gaaaggaacg tgaaaggcat
     1201 gtttgaggtg ctggagccct tgcatgctat gatggaacgg ggcccccaga ctctgaagga
     1261 aacatccttt aatcaggcct atggtcgaga tttaatggag gcccaagagt ggtgcaggaa
     1321 gtacatgaaa tcagggaatg tcaaggacct cacccaagcc tgggacctct attatcatgt
     1381 gttccgacga atctcataag aattcagatc cgcccctctc cctccccccc ccctaacgtt
     1441 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     1501 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     1561 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     1621 gaagcagttc ctctggtagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     1681 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     1741 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     1801 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     1861 cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg
     1921 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     1981 tgataatatg gccacaacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     2041 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     2101 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     2161 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     2221 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     2281 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     2341 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     2401 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     2461 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     2521 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     2581 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     2641 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     2701 gtacaagtaa accggtgctc gaggtaccga tgaatagcta aggtcgaggc cgcaggtaag
     2761 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
     2821 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     2881 ctccacaggt gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2941 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3001 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3061 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3121 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3181 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3241 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     3301 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3361 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3421 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3481 cccgcctgga attcgagctc ggtacgatca gcctcgactg tgccttctag ttgccagcca
     3541 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3601 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3661 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag cccagctttt
     3721 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3781 tgtgaaattg ttatccgcta attcactcct caggtgcagg ctgcctatca gaaggtggtg
     3841 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     3901 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     3961 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     4021 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     4081 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     4141 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     4201 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     4261 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     4321 tggagatcgg ccggccgccg gcgtcgacgc gcgcggccgc ggcgcgccgt tgctggcgtt
     4381 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4441 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4501 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4561 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4621 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4681 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4741 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4801 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4861 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4921 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4981 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5041 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5101 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5161 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5221 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5281 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5341 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5401 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5461 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5521 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5581 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5641 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5701 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5761 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5821 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5881 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5941 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6001 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6061 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6181 tatcacgagg ccctttcgtc tacgcgcgcg ccggcggcgc gccgtcgacg gccggccgcg
     6241 gccgc
//
