LOCUS       sequence-270541-        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-539    C-Tag Venus210[2] L32 Itgb1 mCherry
FEATURES             Location/Qualifiers
     source          1..8450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    join(3617..4191,4192..4194,4195..4203)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3784..3801)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4011..4030
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4204..4914
                     /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(4349..4367)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4617..4636
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4960..5092
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5101..5689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5154..5174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5572..5583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5716..5733)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5722..5833
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5929..5949)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5929..5947)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5968..5984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5968..5984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5981..6003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6086..6105)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6151..6206
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6205..6224)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6585..7173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6665..6684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7344..8135,8136..8204))
                     /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     7967..7986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8205..8309)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8377..8395
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctgcctcca ggcctcaata ttggccatta gccatattat tcattggtta tatagcataa
       61 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      121 attggctcat gtccaatatg accgccatgt tggcattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggcc
     1021 accatgaatt tgcaactggt ttcctggatt ggattgatca gtttgatttg ttctgtattt
     1081 ggccaaacag ataaaaatag atgtttaaaa gcaaatgcca aatcttgcgg agaatgtata
     1141 caagcagggc caaattgtgg gtggtgtaca aatacgacat ttttgcaaga aggaatgcct
     1201 acttctgcac gatgtgatga tttagaagct ttgaaaaaga agggttgcca gccaagtgac
     1261 atagagaatc ccagaggctc tcaaactata aagaaaaata aaaatgtcac caatcgcagc
     1321 aaagggatgg cagagaagct ccggccagaa gacattactc agatccaacc acaacagctg
     1381 cttctaaaat tgagatcagg agaaccacag aagtttacat taaaattcaa gagggctgaa
     1441 gattacccta ttgatctcta ctaccttatg gatctctcct actctatgaa agatgatctg
     1501 gagaatgtga aaagtcttgg aacggatttg atgaatgaaa tgaggaggat tacttcagac
     1561 ttccgcattg gctttggctc atttgtggag aaaactgtga tgccgtatat tagcacaacc
     1621 ccagcaaagc taagaaatcc ttgtacaagt gaacaaaact gcaccagccc atttagctac
     1681 aaaaatgtgc ttagtcttac tgacagagga gagtttttca atgaacttgt tggtcagcaa
     1741 cgcatatctg gaaacttgga ttctccagaa ggtggctttg atgcaatcat gcaggttgcg
     1801 gtttgtggat cgctgattgg ctggaggaat gtaacacgac tgctggtgtt ttccacggat
     1861 gctgggtttc actttgctgg agatgggaaa cttggtggta ttgttttacc caatgatgga
     1921 caatgtcacc tggaaaataa tgtatataca atgagccatt actatgatta tccttcaatt
     1981 gctcaccttg ttcagaaact aagtgaaaat aatattcaga cgatttttgc agttactgaa
     2041 gagttccaac ctgtttacaa ggaattgaag aatttgattc ctaagtcagc agtgggcaca
     2101 ctgtctggaa actctagtaa tgtgatccag ctaatcatcg atgcctacaa ctctctttct
     2161 tcagaagtca ttctggaaaa tagcaaattg ccagacggag taacaataaa ttacaaatcc
     2221 tattgcaaga atggggtgaa tgggacagga gaaaatggac gaaagtgttc caacatttct
     2281 attggagatg aggttcaatt tgaaattagc ataactgcta ataaatgtcc aaataaggag
     2341 tctgaaacca ttaaaattaa acctctgggc ttcactgaag aagtagaggt cgttcttcag
     2401 ttcatctgta agtgcaattg tcaaagccat ggcatcccag ccagtcccaa gtgccatgag
     2461 ggaaatggga catttgagtg tggagcctgc aggtgcaatg aggggcgtgt tgggaggcac
     2521 tgtgaatgta gcacagatga agtgaacagt gaagacatgg acgcttactg caggaaagag
     2581 aacagttcgg aaatctgcag taataatgga gaatgtgtct gtggacagtg tgtgtgtagg
     2641 aagagagata atacaaatga aatttactct ggaaaattct gcgagtgtga taacttcaac
     2701 tgtgataggt ctaatggctt aatttgtgga ggcaatggcg tgtgcaggtg tcgtgtttgt
     2761 gaatgctatc ccaattacac tggcagtgca tgtgactgtt ctttggacac tggtccatgt
     2821 ctagcgtcaa atggtcagat ctgcaatggc cggggtattt gtgaatgtgg tgcttgtaag
     2881 tgcacagatc ccaagtttca agggccaact tgtgagacat gtcagacctg ccttggcgtc
     2941 tgtgcagagc ataaagaatg tgttcagtgc agagccttca ataaaggaga aaagaaagac
     3001 acgtgtgcac aggagtgctc ccacttcaat ctcaccaaag tagaaagcag ggagaagttg
     3061 ccccagccgg tgcaggtcga tcctgtgacc cattgcaagg agaaggacat tgatgactgc
     3121 tggttctatt tcacctattc agtgaatggc aacaatgaag ctatcgtgca tgttgtggag
     3181 actccagact gtcctactgg tcccgacatc atcccaattg tagcaggcgt ggttgctgga
     3241 attgttctta ttggccttgc cttgctgctg atttggaaac ttttaatgat aattcatgac
     3301 agaagggaat ttgctaaatt tgaaaaggag aaaatgaatg ccaagtggga cacgggtgaa
     3361 aatcctattt acaagagtgc cgtgacaact gtggtcaatc cgaagtatga gggaaaagac
     3421 gtgggtggcg gagggagcga aggcggcgga tccggagggc ctggctctgg cggagagggg
     3481 agcgctggag gcgggtcagc cggtggtggc agtgatccca atgagaagag ggaccatatg
     3541 gtactgctcg aattcgtgac tgcagcaggc attacccttg ggatggatga actttataag
     3601 taagaattca gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     3661 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     3721 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     3781 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     3841 tagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     3901 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     3961 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     4021 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     4081 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     4141 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     4201 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     4261 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     4321 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     4381 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     4441 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     4501 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     4561 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     4621 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     4681 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4741 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4801 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     4861 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaaccggt
     4921 gctcgaggta ccgatgaata gctaaggtcg aggccgcagg taagtatcaa ggttacaaga
     4981 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     5041 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtcgac
     5101 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5161 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5221 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5281 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5341 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5401 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5461 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     5521 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5581 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5641 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
     5701 gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5761 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5821 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5881 aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc tttagtgagg
     5941 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6001 gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     6061 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     6121 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     6181 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     6241 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     6301 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     6361 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     6421 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     6481 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tcggccggcc
     6541 gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca taggctccgc
     6601 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6661 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6721 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6781 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6841 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6901 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6961 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7021 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7201 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7261 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7321 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7381 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7441 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7501 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7561 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7621 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7681 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7741 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7801 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7861 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7921 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7981 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8041 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8101 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8161 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8221 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8281 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8341 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     8401 cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc cgcggccgcg
//
