LOCUS       sequence-249253-        6084 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 6084)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6084)
  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-372 N-Tag Venus210[2] L32 {Esp3I(2x)} mC...
FEATURES             Location/Qualifiers
     source          1..6084
                     /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"
     misc_feature    join(1252..1826,1827..1829,1830..1838)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1419..1436)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1646..1665
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1839..2549
                     /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(1984..2002)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2252..2271
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2595..2727
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2736..3324
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2789..2809)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3207..3218)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3351..3368)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3357..3468
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3564..3584)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3564..3582)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3603..3619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3603..3619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3616..3638)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3721..3740)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3786..3841
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3840..3859)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4220..4808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4300..4319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4979..5770,5771..5839))
                     /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     5602..5621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5840..5944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6012..6030
                     /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 cccaatgaga agagggacca tatggtactg ctcgaattcg tgactgcagc
     1081 aggcattacc cttgggatgg atgaacttta taaggacgtg ggtggcggag ggagcgaagg
     1141 cggcggatcc ggagggcctg gctctggcgg agaggggagc gctggaggcg ggtcagccgg
     1201 tggtgggagt cgagacgccg aggcacgccg tctcctaaga attcagatcc gcccctctcc
     1261 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     1321 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     1381 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     1441 tctgttgaat gtcgtgaagg aagcagttcc tctggtagct tcttgaagac aaacaacgtc
     1501 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     1561 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     1621 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     1681 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt
     1741 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     1801 tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagg
     1861 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1921 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1981 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     2041 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     2101 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2161 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     2221 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     2281 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     2341 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2401 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2461 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2521 ccggcggcat ggacgagctg tacaagtaaa ccggtgctcg aggtaccgat gaatagctaa
     2581 ggtcgaggcc gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     2641 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     2701 acatccactt tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt
     2761 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2821 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2881 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2941 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3001 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3061 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3121 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     3181 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3241 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3301 tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt
     3361 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3421 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3481 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3541 agacaatagc ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     3601 tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc aggtgcaggc
     3661 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     3721 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     3781 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     3841 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     3901 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     3961 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     4021 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     4081 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     4141 agctgtccct cttctcttat ggagatcggc cggccgccgg cgtcgacgcg cgcggccgcg
     4201 gcgcgccgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4261 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4321 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4381 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4441 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4501 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4561 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4621 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4681 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4741 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4801 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4861 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5941 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6001 taacctataa aaataggcgt atcacgaggc cctttcgtct acgcgcgcgc cggcggcgcg
     6061 ccgtcgacgg ccggccgcgg ccgc
//
