LOCUS       sequence-249397-        8450 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 8450)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8450)
  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-540    N-Tag Venus210[2] L32 Itgb1 mCherry
FEATURES             Location/Qualifiers
     source          1..8450
                     /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(3618..4192,4193..4195,4196..4204)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3785..3802)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4012..4031
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4205..4915
                     /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(4350..4368)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4618..4637
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4961..5093
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5102..5690
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5155..5175)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5573..5584)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5717..5734)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5723..5834
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5930..5950)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5930..5948)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5969..5985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5969..5985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5982..6004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6087..6106)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6152..6207
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6206..6225)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6586..7174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6666..6685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7345..8136,8137..8205))
                     /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     7968..7987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8206..8310)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8378..8396
                     /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 aatttgcaac tggtttcctg gattggattg atcagtttga tttgttctgt
     1261 atttggccaa acagataaaa atagatgttt aaaagcaaat gccaaatctt gcggagaatg
     1321 tatacaagca gggccaaatt gtgggtggtg tacaaatacg acatttttgc aagaaggaat
     1381 gcctacttct gcacgatgtg atgatttaga agctttgaaa aagaagggtt gccagccaag
     1441 tgacatagag aatcccagag gctctcaaac tataaagaaa aataaaaatg tcaccaatcg
     1501 cagcaaaggg atggcagaga agctccggcc agaagacatt actcagatcc aaccacaaca
     1561 gctgcttcta aaattgagat caggagaacc acagaagttt acattaaaat tcaagagggc
     1621 tgaagattac cctattgatc tctactacct tatggatctc tcctactcta tgaaagatga
     1681 tctggagaat gtgaaaagtc ttggaacgga tttgatgaat gaaatgagga ggattacttc
     1741 agacttccgc attggctttg gctcatttgt ggagaaaact gtgatgccgt atattagcac
     1801 aaccccagca aagctaagaa atccttgtac aagtgaacaa aactgcacca gcccatttag
     1861 ctacaaaaat gtgcttagtc ttactgacag aggagagttt ttcaatgaac ttgttggtca
     1921 gcaacgcata tctggaaact tggattctcc agaaggtggc tttgatgcaa tcatgcaggt
     1981 tgcggtttgt ggatcgctga ttggctggag gaatgtaaca cgactgctgg tgttttccac
     2041 ggatgctggg tttcactttg ctggagatgg gaaacttggt ggtattgttt tacccaatga
     2101 tggacaatgt cacctggaaa ataatgtata tacaatgagc cattactatg attatccttc
     2161 aattgctcac cttgttcaga aactaagtga aaataatatt cagacgattt ttgcagttac
     2221 tgaagagttc caacctgttt acaaggaatt gaagaatttg attcctaagt cagcagtggg
     2281 cacactgtct ggaaactcta gtaatgtgat ccagctaatc atcgatgcct acaactctct
     2341 ttcttcagaa gtcattctgg aaaatagcaa attgccagac ggagtaacaa taaattacaa
     2401 atcctattgc aagaatgggg tgaatgggac aggagaaaat ggacgaaagt gttccaacat
     2461 ttctattgga gatgaggttc aatttgaaat tagcataact gctaataaat gtccaaataa
     2521 ggagtctgaa accattaaaa ttaaacctct gggcttcact gaagaagtag aggtcgttct
     2581 tcagttcatc tgtaagtgca attgtcaaag ccatggcatc ccagccagtc ccaagtgcca
     2641 tgagggaaat gggacatttg agtgtggagc ctgcaggtgc aatgaggggc gtgttgggag
     2701 gcactgtgaa tgtagcacag atgaagtgaa cagtgaagac atggacgctt actgcaggaa
     2761 agagaacagt tcggaaatct gcagtaataa tggagaatgt gtctgtggac agtgtgtgtg
     2821 taggaagaga gataatacaa atgaaattta ctctggaaaa ttctgcgagt gtgataactt
     2881 caactgtgat aggtctaatg gcttaatttg tggaggcaat ggcgtgtgca ggtgtcgtgt
     2941 ttgtgaatgc tatcccaatt acactggcag tgcatgtgac tgttctttgg acactggtcc
     3001 atgtctagcg tcaaatggtc agatctgcaa tggccggggt atttgtgaat gtggtgcttg
     3061 taagtgcaca gatcccaagt ttcaagggcc aacttgtgag acatgtcaga cctgccttgg
     3121 cgtctgtgca gagcataaag aatgtgttca gtgcagagcc ttcaataaag gagaaaagaa
     3181 agacacgtgt gcacaggagt gctcccactt caatctcacc aaagtagaaa gcagggagaa
     3241 gttgccccag ccggtgcagg tcgatcctgt gacccattgc aaggagaagg acattgatga
     3301 ctgctggttc tatttcacct attcagtgaa tggcaacaat gaagctatcg tgcatgttgt
     3361 ggagactcca gactgtccta ctggtcccga catcatccca attgtagcag gcgtggttgc
     3421 tggaattgtt cttattggcc ttgccttgct gctgatttgg aaacttttaa tgataattca
     3481 tgacagaagg gaatttgcta aatttgaaaa ggagaaaatg aatgccaagt gggacacggg
     3541 tgaaaatcct atttacaaga gtgccgtgac aactgtggtc aatccgaagt atgagggaaa
     3601 ataagaattc agatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     3661 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     3721 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     3781 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     3841 gtagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     3901 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     3961 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     4021 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     4081 gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     4141 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     4201 aaccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     4261 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     4321 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     4381 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     4441 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     4501 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     4561 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     4621 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     4681 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     4741 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     4801 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     4861 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaaccgg
     4921 tgctcgaggt accgatgaat agctaaggtc gaggccgcag gtaagtatca aggttacaag
     4981 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
     5041 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcga
     5101 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5161 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5221 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5281 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5341 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5401 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5461 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     5521 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5581 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5641 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg
     5701 agctcggtac gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     5761 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     5821 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     5881 caggacagca agggggagga ttgggaagac aatagcccag cttttgttcc ctttagtgag
     5941 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6001 cgctaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     6061 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     6121 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     6181 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     6241 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     6301 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     6361 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     6421 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     6481 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atcggccggc
     6541 cgccggcgtc gacgcgcgcg gccgcggcgc gccgttgctg gcgtttttcc ataggctccg
     6601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7141 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7201 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7261 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7321 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7501 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7621 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8221 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8281 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8341 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     8401 tcgtctacgc gcgcgccggc ggcgcgccgt cgacggccgg ccgcggccgc
//
