LOCUS       sequence-308459-        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"
     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(3581..4155,4156..4158,4159..4167)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3748..3765)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3975..3994
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4168..4878
                     /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(4313..4331)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4581..4600
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4924..5056
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5065..5653
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5118..5138)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5536..5547)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5680..5697)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5686..5797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5893..5913)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5893..5911)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5932..5948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5932..5948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5945..5967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6050..6069)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6115..6170
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6169..6188)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6549..7137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6629..6648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7308..8099,8100..8168))
                     /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     7931..7950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8169..8273)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8341..8359
                     /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 gggaaaaggg
     3421 agtcctggcg gtggatccgg aggctctggg agctctgcca gcggtggcag caccagtgat
     3481 cccaatgaga agagggacca tatggtactg ctcgaattcg tgactgcagc aggcattacc
     3541 cttgggatgg atgaacttta taagtaagaa ttcagatccg cccctctccc tccccccccc
     3601 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     3661 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     3721 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     3781 tcgtgaagga agcagttcct ctggtagctt cttgaagaca aacaacgtct gtagcgaccc
     3841 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     3901 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     3961 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     4021 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
     4081 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     4141 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagga taacatggcc
     4201 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     4261 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     4321 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     4381 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     4441 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     4501 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     4561 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     4621 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     4681 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     4741 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     4801 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     4861 gacgagctgt acaagtaaac cggtgctcga ggtaccgatg aatagctaag gtcgaggccg
     4921 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
     4981 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     5041 gcctttctct ccacaggtgt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     5101 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5161 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5221 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5281 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5341 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     5401 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     5461 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5521 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5581 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5641 gccgcctccc cgcctggaat tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt
     5701 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5761 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5821 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagcc
     5881 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     5941 gtttcctgtg tgaaattgtt atccgctaat tcactcctca ggtgcaggct gcctatcaga
     6001 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     6061 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     6121 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     6181 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     6241 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     6301 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     6361 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     6421 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     6481 ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc gcggccgcgg cgcgccgttg
     6541 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6601 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6661 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6721 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6781 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6841 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6901 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6961 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7021 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7081 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7141 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7201 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7441 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8281 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     8341 aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc ggcggcgcgc cgtcgacggc
     8401 cggccgcggc cgcg
//
