LOCUS       sequence-249224-        6093 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 6093)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6093)
  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-343    C-Tag UnaG[2] L10 {Esp3I(2x)} mCh...
FEATURES             Location/Qualifiers
     source          1..6093
                     /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"
     misc_feature    join(1261..1835,1836..1838,1839..1847)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1428..1445)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1655..1674
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1848..2558
                     /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(1993..2011)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2261..2280
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2604..2736
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2745..3333
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2798..2818)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3216..3227)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3360..3377)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3366..3477
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3573..3593)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3573..3591)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3612..3628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3612..3628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3625..3647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3730..3749)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3795..3850
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3849..3868)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4229..4817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4309..4328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4988..5779,5780..5848))
                     /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     5611..5630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5849..5953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6021..6039
                     /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 cacccgagac gccgaggcac gccgtctccg ggagctctgc cagcggtggc agcaccagtg
     1081 gggtgaagtc tgtggtgaat ctggtaggcg agaaactcgt gtacgtccag aagtgggatg
     1141 ggaaggaaac cacgtatgtg cgagagatca aggacgggaa gttggttgtc accctgacca
     1201 tgggcgatgt ggttgctgtg cgctcatatc ggagagccac tgaataagaa ttcagatccg
     1261 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     1321 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     1381 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     1441 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggtagctt cttgaagaca
     1501 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     1561 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     1621 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     1681 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggta
     1741 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     1801 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     1861 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     1921 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1981 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2041 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     2101 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2161 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     2221 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2281 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     2341 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     2401 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     2461 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2521 gccactccac cggcggcatg gacgagctgt acaagtaaac cggtgctcga ggtaccgatg
     2581 aatagctaag gtcgaggccg caggtaagta tcaaggttac aagacaggtt taaggagacc
     2641 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     2701 gtcttactga catccacttt gcctttctct ccacaggtgt cgacaatcaa cctctggatt
     2761 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2821 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2881 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2941 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3001 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3061 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3121 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3181 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3241 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3301 cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacgatcagc
     3361 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3421 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3481 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3541 ggattgggaa gacaatagcc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     3601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctaat tcactcctca
     3661 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     3721 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     3781 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3841 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     3901 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     3961 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     4021 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     4081 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     4141 cccagtcata gctgtccctc ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc
     4201 gcggccgcgg cgcgccgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6001 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc
     6061 ggcggcgcgc cgtcgacggc cggccgcggc cgc
//
