LOCUS       sequence-249419-        8384 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 8384)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8384)
  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-514    N-Tag GFP[1] L20 Itgb1 mCherry
FEATURES             Location/Qualifiers
     source          1..8384
                     /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(3552..4126,4127..4129,4130..4138)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3719..3736)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3946..3965
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4139..4849
                     /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(4284..4302)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4552..4571
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4895..5027
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5036..5624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5089..5109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5507..5518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5651..5668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5657..5768
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5864..5884)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5864..5882)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5903..5919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5903..5919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5916..5938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6021..6040)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6086..6141
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6140..6159)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6520..7108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6600..6619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7279..8070,8071..8139))
                     /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     7902..7921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8140..8244)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8312..8330
                     /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 taacgggagt cctggcggtg gatccggagg ctctgggagc tctgccagcg gtggcagcac
     1141 ctccaatttg caactggttt cctggattgg attgatcagt ttgatttgtt ctgtatttgg
     1201 ccaaacagat aaaaatagat gtttaaaagc aaatgccaaa tcttgcggag aatgtataca
     1261 agcagggcca aattgtgggt ggtgtacaaa tacgacattt ttgcaagaag gaatgcctac
     1321 ttctgcacga tgtgatgatt tagaagcttt gaaaaagaag ggttgccagc caagtgacat
     1381 agagaatccc agaggctctc aaactataaa gaaaaataaa aatgtcacca atcgcagcaa
     1441 agggatggca gagaagctcc ggccagaaga cattactcag atccaaccac aacagctgct
     1501 tctaaaattg agatcaggag aaccacagaa gtttacatta aaattcaaga gggctgaaga
     1561 ttaccctatt gatctctact accttatgga tctctcctac tctatgaaag atgatctgga
     1621 gaatgtgaaa agtcttggaa cggatttgat gaatgaaatg aggaggatta cttcagactt
     1681 ccgcattggc tttggctcat ttgtggagaa aactgtgatg ccgtatatta gcacaacccc
     1741 agcaaagcta agaaatcctt gtacaagtga acaaaactgc accagcccat ttagctacaa
     1801 aaatgtgctt agtcttactg acagaggaga gtttttcaat gaacttgttg gtcagcaacg
     1861 catatctgga aacttggatt ctccagaagg tggctttgat gcaatcatgc aggttgcggt
     1921 ttgtggatcg ctgattggct ggaggaatgt aacacgactg ctggtgtttt ccacggatgc
     1981 tgggtttcac tttgctggag atgggaaact tggtggtatt gttttaccca atgatggaca
     2041 atgtcacctg gaaaataatg tatatacaat gagccattac tatgattatc cttcaattgc
     2101 tcaccttgtt cagaaactaa gtgaaaataa tattcagacg atttttgcag ttactgaaga
     2161 gttccaacct gtttacaagg aattgaagaa tttgattcct aagtcagcag tgggcacact
     2221 gtctggaaac tctagtaatg tgatccagct aatcatcgat gcctacaact ctctttcttc
     2281 agaagtcatt ctggaaaata gcaaattgcc agacggagta acaataaatt acaaatccta
     2341 ttgcaagaat ggggtgaatg ggacaggaga aaatggacga aagtgttcca acatttctat
     2401 tggagatgag gttcaatttg aaattagcat aactgctaat aaatgtccaa ataaggagtc
     2461 tgaaaccatt aaaattaaac ctctgggctt cactgaagaa gtagaggtcg ttcttcagtt
     2521 catctgtaag tgcaattgtc aaagccatgg catcccagcc agtcccaagt gccatgaggg
     2581 aaatgggaca tttgagtgtg gagcctgcag gtgcaatgag gggcgtgttg ggaggcactg
     2641 tgaatgtagc acagatgaag tgaacagtga agacatggac gcttactgca ggaaagagaa
     2701 cagttcggaa atctgcagta ataatggaga atgtgtctgt ggacagtgtg tgtgtaggaa
     2761 gagagataat acaaatgaaa tttactctgg aaaattctgc gagtgtgata acttcaactg
     2821 tgataggtct aatggcttaa tttgtggagg caatggcgtg tgcaggtgtc gtgtttgtga
     2881 atgctatccc aattacactg gcagtgcatg tgactgttct ttggacactg gtccatgtct
     2941 agcgtcaaat ggtcagatct gcaatggccg gggtatttgt gaatgtggtg cttgtaagtg
     3001 cacagatccc aagtttcaag ggccaacttg tgagacatgt cagacctgcc ttggcgtctg
     3061 tgcagagcat aaagaatgtg ttcagtgcag agccttcaat aaaggagaaa agaaagacac
     3121 gtgtgcacag gagtgctccc acttcaatct caccaaagta gaaagcaggg agaagttgcc
     3181 ccagccggtg caggtcgatc ctgtgaccca ttgcaaggag aaggacattg atgactgctg
     3241 gttctatttc acctattcag tgaatggcaa caatgaagct atcgtgcatg ttgtggagac
     3301 tccagactgt cctactggtc ccgacatcat cccaattgta gcaggcgtgg ttgctggaat
     3361 tgttcttatt ggccttgcct tgctgctgat ttggaaactt ttaatgataa ttcatgacag
     3421 aagggaattt gctaaatttg aaaaggagaa aatgaatgcc aagtgggaca cgggtgaaaa
     3481 tcctatttac aagagtgccg tgacaactgt ggtcaatccg aagtatgagg gaaaataaga
     3541 attcagatcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     3601 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3661 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3721 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggtagct
     3781 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     3841 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     3901 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     3961 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     4021 gggcctcggt acacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     4081 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     4141 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     4201 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     4261 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     4321 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     4381 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     4441 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     4501 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     4561 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     4621 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     4681 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     4741 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     4801 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaaa ccggtgctcg
     4861 aggtaccgat gaatagctaa ggtcgaggcc gcaggtaagt atcaaggtta caagacaggt
     4921 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     4981 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tcgacaatca
     5041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5161 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5221 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5281 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5341 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5401 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     5461 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5521 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     5581 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg
     5641 gtacgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     5701 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     5761 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     5821 agcaaggggg aggattggga agacaatagc ccagcttttg ttccctttag tgagggttaa
     5881 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa
     5941 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     6001 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     6061 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     6121 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     6181 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     6241 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     6301 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     6361 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     6421 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatcggc cggccgccgg
     6481 cgtcgacgcg cgcggccgcg gcgcgccgtt gctggcgttt ttccataggc tccgcccccc
     6541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8281 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8341 acgcgcgcgc cggcggcgcg ccgtcgacgg ccggccgcgg ccgc
//
