LOCUS       sequence-249417-        8354 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 8354)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8354)
  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-512    N-Tag GFP[1] L10 Itgb1 mCherry
FEATURES             Location/Qualifiers
     source          1..8354
                     /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(3522..4096,4097..4099,4100..4108)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3689..3706)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3916..3935
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4109..4819
                     /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(4254..4272)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4522..4541
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4865..4997
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5006..5594
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5059..5079)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5477..5488)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5621..5638)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5627..5738
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5834..5854)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5834..5852)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5873..5889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5873..5889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5886..5908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5991..6010)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6056..6111
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6110..6129)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6490..7078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6570..6589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7249..8040,8041..8109))
                     /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     7872..7891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8110..8214)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8282..8300
                     /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 ctgcctgatg accactacct gagcacacag accatcctct ccaaggacct
     1081 taacgggagc tctgccagcg gtggcagcac ctccaatttg caactggttt cctggattgg
     1141 attgatcagt ttgatttgtt ctgtatttgg ccaaacagat aaaaatagat gtttaaaagc
     1201 aaatgccaaa tcttgcggag aatgtataca agcagggcca aattgtgggt ggtgtacaaa
     1261 tacgacattt ttgcaagaag gaatgcctac ttctgcacga tgtgatgatt tagaagcttt
     1321 gaaaaagaag ggttgccagc caagtgacat agagaatccc agaggctctc aaactataaa
     1381 gaaaaataaa aatgtcacca atcgcagcaa agggatggca gagaagctcc ggccagaaga
     1441 cattactcag atccaaccac aacagctgct tctaaaattg agatcaggag aaccacagaa
     1501 gtttacatta aaattcaaga gggctgaaga ttaccctatt gatctctact accttatgga
     1561 tctctcctac tctatgaaag atgatctgga gaatgtgaaa agtcttggaa cggatttgat
     1621 gaatgaaatg aggaggatta cttcagactt ccgcattggc tttggctcat ttgtggagaa
     1681 aactgtgatg ccgtatatta gcacaacccc agcaaagcta agaaatcctt gtacaagtga
     1741 acaaaactgc accagcccat ttagctacaa aaatgtgctt agtcttactg acagaggaga
     1801 gtttttcaat gaacttgttg gtcagcaacg catatctgga aacttggatt ctccagaagg
     1861 tggctttgat gcaatcatgc aggttgcggt ttgtggatcg ctgattggct ggaggaatgt
     1921 aacacgactg ctggtgtttt ccacggatgc tgggtttcac tttgctggag atgggaaact
     1981 tggtggtatt gttttaccca atgatggaca atgtcacctg gaaaataatg tatatacaat
     2041 gagccattac tatgattatc cttcaattgc tcaccttgtt cagaaactaa gtgaaaataa
     2101 tattcagacg atttttgcag ttactgaaga gttccaacct gtttacaagg aattgaagaa
     2161 tttgattcct aagtcagcag tgggcacact gtctggaaac tctagtaatg tgatccagct
     2221 aatcatcgat gcctacaact ctctttcttc agaagtcatt ctggaaaata gcaaattgcc
     2281 agacggagta acaataaatt acaaatccta ttgcaagaat ggggtgaatg ggacaggaga
     2341 aaatggacga aagtgttcca acatttctat tggagatgag gttcaatttg aaattagcat
     2401 aactgctaat aaatgtccaa ataaggagtc tgaaaccatt aaaattaaac ctctgggctt
     2461 cactgaagaa gtagaggtcg ttcttcagtt catctgtaag tgcaattgtc aaagccatgg
     2521 catcccagcc agtcccaagt gccatgaggg aaatgggaca tttgagtgtg gagcctgcag
     2581 gtgcaatgag gggcgtgttg ggaggcactg tgaatgtagc acagatgaag tgaacagtga
     2641 agacatggac gcttactgca ggaaagagaa cagttcggaa atctgcagta ataatggaga
     2701 atgtgtctgt ggacagtgtg tgtgtaggaa gagagataat acaaatgaaa tttactctgg
     2761 aaaattctgc gagtgtgata acttcaactg tgataggtct aatggcttaa tttgtggagg
     2821 caatggcgtg tgcaggtgtc gtgtttgtga atgctatccc aattacactg gcagtgcatg
     2881 tgactgttct ttggacactg gtccatgtct agcgtcaaat ggtcagatct gcaatggccg
     2941 gggtatttgt gaatgtggtg cttgtaagtg cacagatccc aagtttcaag ggccaacttg
     3001 tgagacatgt cagacctgcc ttggcgtctg tgcagagcat aaagaatgtg ttcagtgcag
     3061 agccttcaat aaaggagaaa agaaagacac gtgtgcacag gagtgctccc acttcaatct
     3121 caccaaagta gaaagcaggg agaagttgcc ccagccggtg caggtcgatc ctgtgaccca
     3181 ttgcaaggag aaggacattg atgactgctg gttctatttc acctattcag tgaatggcaa
     3241 caatgaagct atcgtgcatg ttgtggagac tccagactgt cctactggtc ccgacatcat
     3301 cccaattgta gcaggcgtgg ttgctggaat tgttcttatt ggccttgcct tgctgctgat
     3361 ttggaaactt ttaatgataa ttcatgacag aagggaattt gctaaatttg aaaaggagaa
     3421 aatgaatgcc aagtgggaca cgggtgaaaa tcctatttac aagagtgccg tgacaactgt
     3481 ggtcaatccg aagtatgagg gaaaataaga attcagatcc gcccctctcc ctcccccccc
     3541 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     3601 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     3661 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     3721 gtcgtgaagg aagcagttcc tctggtagct tcttgaagac aaacaacgtc tgtagcgacc
     3781 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     3841 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     3901 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     3961 aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt
     4021 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     4081 aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagg ataacatggc
     4141 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     4201 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4261 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     4321 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     4381 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     4441 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     4501 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     4561 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     4621 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     4681 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     4741 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     4801 ggacgagctg tacaagtaaa ccggtgctcg aggtaccgat gaatagctaa ggtcgaggcc
     4861 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
     4921 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     4981 tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5041 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5101 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5161 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5221 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5281 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5341 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5401 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5461 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5521 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5581 ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt gccttctagt
     5641 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5701 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5761 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     5821 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     5881 tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc aggtgcaggc tgcctatcag
     5941 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     6001 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     6061 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     6121 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     6181 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     6241 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     6301 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     6361 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     6421 cttctcttat ggagatcggc cggccgccgg cgtcgacgcg cgcggccgcg gcgcgccgtt
     6481 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6541 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6601 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6661 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6721 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6781 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6841 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6901 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6961 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7021 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7081 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7141 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7201 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7261 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7321 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7381 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7441 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7501 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7561 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7621 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7681 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7741 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7801 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7861 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7921 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7981 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8041 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8101 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8161 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8221 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8281 aaataggcgt atcacgaggc cctttcgtct acgcgcgcgc cggcggcgcg ccgtcgacgg
     8341 ccggccgcgg ccgc
//
