LOCUS       sequence-249238-        6210 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 6210)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6210)
  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-357 C-Tag Venus173/155[2] L20 {Esp3I(2x)...
FEATURES             Location/Qualifiers
     source          1..6210
                     /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"
     misc_feature    join(1378..1952,1953..1955,1956..1964)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1545..1562)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1772..1791
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1965..2675
                     /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(2110..2128)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2378..2397
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2721..2853
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2862..3450
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2915..2935)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3333..3344)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3477..3494)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3483..3594
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3690..3710)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3690..3708)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3729..3745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3729..3745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3742..3764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3847..3866)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3912..3967
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3966..3985)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4346..4934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4426..4445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5105..5896,5897..5965))
                     /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     5728..5747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5966..6070)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6138..6156
                     /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 cacccgagac gccgaggcac gccgtctccg ggagtcctgg cggtggatcc ggaggctctg
     1081 ggagctctgc cagcggtggc agcaccagtg ataagcagaa gaatgggatt aaggcgaact
     1141 tcaagatccg acacaacatc gaggatggtg gggtccagct tgccgaccat tatcagcaga
     1201 acacacccat tggggatgga ccagtcctgt tgccggataa ccactacctc agttaccaga
     1261 gcaaactgtc caaagatccc aatgagaaga gggaccatat ggtactgctc gaattcgtga
     1321 ctgcagcagg cattaccctt gggatggatg aactttataa gtaagaattc agatccgccc
     1381 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     1441 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     1501 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     1561 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt gaagacaaac
     1621 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     1681 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     1741 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     1801 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac
     1861 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     1921 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg
     1981 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     2041 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2101 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     2161 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2221 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2281 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2341 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     2401 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     2461 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2521 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2581 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2641 actccaccgg cggcatggac gagctgtaca agtaaaccgg tgctcgaggt accgatgaat
     2701 agctaaggtc gaggccgcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     2761 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     2821 ttactgacat ccactttgcc tttctctcca caggtgtcga caatcaacct ctggattaca
     2881 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2941 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3001 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3061 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3121 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3181 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3241 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3301 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3361 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3421 gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac gatcagcctc
     3481 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3541 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3601 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3661 ttgggaagac aatagcccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     3721 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctaattca ctcctcaggt
     3781 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     3841 ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     3901 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3961 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     4021 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     4081 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     4141 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     4201 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     4261 agtcatagct gtccctcttc tcttatggag atcggccggc cgccggcgtc gacgcgcgcg
     4321 gccgcggcgc gccgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4981 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5041 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5101 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5161 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5221 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5281 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5341 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5401 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5461 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5521 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5581 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5641 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5701 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5761 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5821 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5881 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5941 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6001 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6061 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6121 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctacgc gcgcgccggc
     6181 ggcgcgccgt cgacggccgg ccgcggccgc
//
