LOCUS       sequence-270540-        8414 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 8414)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8414)
  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-537    C-Tag Venus210[2] L20 Itgb1 mCherry
FEATURES             Location/Qualifiers
     source          1..8414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(419..993,994..996,997..1005)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(586..603)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     813..832
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1006..1716
                     /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(1151..1169)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1419..1438
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          1762..1894
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    1903..2491
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1956..1976)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2374..2385)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2518..2535)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2524..2635
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2731..2751)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2731..2749)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2770..2786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2770..2786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2783..2805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(2888..2907)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2953..3008
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3007..3026)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(3387..3975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3467..3486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4146..4937,4938..5006))
                     /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     4769..4788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5007..5111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5179..5197
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        5404..5783
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5784..5987
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5938..5958
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5984..6008
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6123..6255
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
ORIGIN
        1 atcgtgcatg ttgtggagac tccagactgt cctactggtc ccgacatcat cccaattgta
       61 gcaggcgtgg ttgctggaat tgttcttatt ggccttgcct tgctgctgat ttggaaactt
      121 ttaatgataa ttcatgacag aagggaattt gctaaatttg aaaaggagaa aatgaatgcc
      181 aagtgggaca cgggtgaaaa tcctatttac aagagtgccg tgacaactgt ggtcaatccg
      241 aagtatgagg gaaaagggag tcctggcggt ggatccggag gctctgggag ctctgccagc
      301 ggtggcagca ccagtgatcc caatgagaag agggaccata tggtactgct cgaattcgtg
      361 actgcagcag gcattaccct tgggatggat gaactttata agtaagaatt cagatccgcc
      421 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
      481 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
      541 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
      601 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggtagcttct tgaagacaaa
      661 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
      721 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
      781 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
      841 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca
      901 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
      961 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc
     1021 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     1081 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1141 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     1201 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     1261 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1321 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     1381 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     1441 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     1501 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     1561 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     1621 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     1681 cactccaccg gcggcatgga cgagctgtac aagtaaaccg gtgctcgagg taccgatgaa
     1741 tagctaaggt cgaggccgca ggtaagtatc aaggttacaa gacaggttta aggagaccaa
     1801 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
     1861 cttactgaca tccactttgc ctttctctcc acaggtgtcg acaatcaacc tctggattac
     1921 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1981 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2041 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2101 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2161 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     2221 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2281 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     2341 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     2401 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     2461 agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta cgatcagcct
     2521 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2581 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2641 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2701 attgggaaga caatagccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     2761 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctaattc actcctcagg
     2821 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     2881 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     2941 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     3001 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     3061 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     3121 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     3181 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     3241 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     3301 cagtcatagc tgtccctctt ctcttatgga gatcggccgg ccgccggcgt cgacgcgcgc
     3361 ggccgcggcg cgccgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4021 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4081 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4261 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4441 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4741 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5161 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctacg cgcgcgccgg
     5221 cggcgcgccg tcgacggccg gccgcggccg cgcctgcctc caggcctcaa tattggccat
     5281 tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     5341 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
     5401 gttggcattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     5461 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5521 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5581 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5641 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
     5701 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
     5761 tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat
     5821 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     5881 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc
     5941 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
     6001 gtcagatcac tagaagcttt attgcggtag tttatcacag ttaaattgct aacgcagtca
     6061 gtgcttctga cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc
     6121 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
     6181 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
     6241 cctttctctc cacaggtgtc cactcccagg ccaccatgaa tttgcaactg gtttcctgga
     6301 ttggattgat cagtttgatt tgttctgtat ttggccaaac agataaaaat agatgtttaa
     6361 aagcaaatgc caaatcttgc ggagaatgta tacaagcagg gccaaattgt gggtggtgta
     6421 caaatacgac atttttgcaa gaaggaatgc ctacttctgc acgatgtgat gatttagaag
     6481 ctttgaaaaa gaagggttgc cagccaagtg acatagagaa tcccagaggc tctcaaacta
     6541 taaagaaaaa taaaaatgtc accaatcgca gcaaagggat ggcagagaag ctccggccag
     6601 aagacattac tcagatccaa ccacaacagc tgcttctaaa attgagatca ggagaaccac
     6661 agaagtttac attaaaattc aagagggctg aagattaccc tattgatctc tactacctta
     6721 tggatctctc ctactctatg aaagatgatc tggagaatgt gaaaagtctt ggaacggatt
     6781 tgatgaatga aatgaggagg attacttcag acttccgcat tggctttggc tcatttgtgg
     6841 agaaaactgt gatgccgtat attagcacaa ccccagcaaa gctaagaaat ccttgtacaa
     6901 gtgaacaaaa ctgcaccagc ccatttagct acaaaaatgt gcttagtctt actgacagag
     6961 gagagttttt caatgaactt gttggtcagc aacgcatatc tggaaacttg gattctccag
     7021 aaggtggctt tgatgcaatc atgcaggttg cggtttgtgg atcgctgatt ggctggagga
     7081 atgtaacacg actgctggtg ttttccacgg atgctgggtt tcactttgct ggagatggga
     7141 aacttggtgg tattgtttta cccaatgatg gacaatgtca cctggaaaat aatgtatata
     7201 caatgagcca ttactatgat tatccttcaa ttgctcacct tgttcagaaa ctaagtgaaa
     7261 ataatattca gacgattttt gcagttactg aagagttcca acctgtttac aaggaattga
     7321 agaatttgat tcctaagtca gcagtgggca cactgtctgg aaactctagt aatgtgatcc
     7381 agctaatcat cgatgcctac aactctcttt cttcagaagt cattctggaa aatagcaaat
     7441 tgccagacgg agtaacaata aattacaaat cctattgcaa gaatggggtg aatgggacag
     7501 gagaaaatgg acgaaagtgt tccaacattt ctattggaga tgaggttcaa tttgaaatta
     7561 gcataactgc taataaatgt ccaaataagg agtctgaaac cattaaaatt aaacctctgg
     7621 gcttcactga agaagtagag gtcgttcttc agttcatctg taagtgcaat tgtcaaagcc
     7681 atggcatccc agccagtccc aagtgccatg agggaaatgg gacatttgag tgtggagcct
     7741 gcaggtgcaa tgaggggcgt gttgggaggc actgtgaatg tagcacagat gaagtgaaca
     7801 gtgaagacat ggacgcttac tgcaggaaag agaacagttc ggaaatctgc agtaataatg
     7861 gagaatgtgt ctgtggacag tgtgtgtgta ggaagagaga taatacaaat gaaatttact
     7921 ctggaaaatt ctgcgagtgt gataacttca actgtgatag gtctaatggc ttaatttgtg
     7981 gaggcaatgg cgtgtgcagg tgtcgtgttt gtgaatgcta tcccaattac actggcagtg
     8041 catgtgactg ttctttggac actggtccat gtctagcgtc aaatggtcag atctgcaatg
     8101 gccggggtat ttgtgaatgt ggtgcttgta agtgcacaga tcccaagttt caagggccaa
     8161 cttgtgagac atgtcagacc tgccttggcg tctgtgcaga gcataaagaa tgtgttcagt
     8221 gcagagcctt caataaagga gaaaagaaag acacgtgtgc acaggagtgc tcccacttca
     8281 atctcaccaa agtagaaagc agggagaagt tgccccagcc ggtgcaggtc gatcctgtga
     8341 cccattgcaa ggagaaggac attgatgact gctggttcta tttcacctat tcagtgaatg
     8401 gcaacaatga agct
//
