LOCUS       sequence-249321-        6440 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 6440)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6440)
  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-284    N-Tag Venus173/155[2] L10 FRB mCh...
FEATURES             Location/Qualifiers
     source          1..6440
                     /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             1316..1591
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1608..2182,2183..2185,2186..2194)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1775..1792)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2002..2021
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2195..2905
                     /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(2340..2358)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2608..2627
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2951..3083
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3092..3680
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3145..3165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3563..3574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3707..3724)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3713..3824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3920..3940)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3920..3938)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3959..3975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3959..3975)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3972..3994)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4077..4096)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4142..4197
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4196..4215)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4576..5164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4656..4675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5335..6126,6127..6195))
                     /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     5958..5977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6196..6300)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6368..6386
                     /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 aagcagaaga atgggattaa ggcgaacttc aagatccgac acaacatcga
     1081 ggatggtggg gtccagcttg ccgaccatta tcagcagaac acacccattg gggatggacc
     1141 agtcctgttg ccggataacc actacctcag ttaccagagc aaactgtcca aagatcccaa
     1201 tgagaagagg gaccatatgg tactgctcga attcgtgact gcagcaggca ttacccttgg
     1261 gatggatgaa ctttataagg ggagctctgc cagcggtggc agcacctccg tggccatcct
     1321 ctggcatgag atgtggcatg aaggcctgga agaggcatct cgtttgtact ttggggaaag
     1381 gaacgtgaaa ggcatgtttg aggtgctgga gcccttgcat gctatgatgg aacggggccc
     1441 ccagactctg aaggaaacat cctttaatca ggcctatggt cgagatttaa tggaggccca
     1501 agagtggtgc aggaagtaca tgaaatcagg gaatgtcaag gacctcaccc aagcctggga
     1561 cctctattat catgtgttcc gacgaatctc ataagaattc agatccgccc ctctccctcc
     1621 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1681 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1741 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1801 ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt gaagacaaac aacgtctgta
     1861 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1921 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     1981 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2041 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     2101 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2161 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggaggataa
     2221 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     2281 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2341 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     2401 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2461 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2521 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     2581 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2641 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     2701 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     2761 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     2821 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     2881 cggcatggac gagctgtaca agtaaaccgg tgctcgaggt accgatgaat agctaaggtc
     2941 gaggccgcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     3001 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
     3061 ccactttgcc tttctctcca caggtgtcga caatcaacct ctggattaca aaatttgtga
     3121 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3181 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3241 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3301 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3361 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3421 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3481 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3541 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3601 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3661 cctttgggcc gcctccccgc ctggaattcg agctcggtac gatcagcctc gactgtgcct
     3721 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3781 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3841 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3901 aatagcccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     3961 catagctgtt tcctgtgtga aattgttatc cgctaattca ctcctcaggt gcaggctgcc
     4021 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     4081 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     4141 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     4201 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     4261 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     4321 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     4381 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     4441 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     4501 gtccctcttc tcttatggag atcggccggc cgccggcgtc gacgcgcgcg gccgcggcgc
     4561 gccgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4621 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4681 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4741 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4801 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4861 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4921 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4981 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5041 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5101 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5161 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5221 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5281 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5341 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5401 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5461 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5521 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5581 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5641 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5701 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5761 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5821 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5881 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5941 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6001 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6061 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6121 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6181 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6241 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6301 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6361 ctataaaaat aggcgtatca cgaggccctt tcgtctacgc gcgcgccggc ggcgcgccgt
     6421 cgacggccgg ccgcggccgc
//
