LOCUS       sequence-249430-        7076 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 7076)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7076)
  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-525    C-Tag GFP[1] L20 Tln1(rod) mCherry
FEATURES             Location/Qualifiers
     source          1..7076
                     /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(2244..2818,2819..2821,2822..2830)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2411..2428)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2638..2657
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2831..3541
                     /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(2976..2994)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3244..3263
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          3587..3719
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3728..4316
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3781..3801)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4199..4210)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4343..4360)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4349..4460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4556..4576)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4556..4574)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4595..4611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4595..4611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4608..4630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4713..4732)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4778..4833
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4832..4851)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5212..5800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5292..5311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5971..6762,6763..6831))
                     /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     6594..6613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6832..6936)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7004..7022
                     /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 caccatggct gtgtctggta tcattgctga cctcgacacc accatcatgt tcgctactgc
     1081 tggcacactt aaccgtgagg gtgctgaaac ttttgctgac caccgggagg gtatcttaaa
     1141 gacagcaaag gttcttgtgg aggacaccaa ggtcctagtg cagaatgcag ctgggagcca
     1201 ggagaagttg gcacaagccg cccagtcctc cgtggccacc attacccgcc tcgctgatgt
     1261 ggtcaagctc ggtgcagcca gcctaggagc cgaagaccct gaaactcagg tggtgctgat
     1321 caatgcagta aaggacgtag ccaaggccct gggtgacctc atcagcgcta cgaaggctgc
     1381 agcgggcaaa gttggggatg accctgcagt gtggcagctc aagaactctg ccaaggtgat
     1441 ggtgaccaat gtgacatcat tgctcaagac agtgaaggct gtggaagatg aggccaccaa
     1501 aggcacacgg gccctagagg caaccacaga gcacatacgt caggaactgg cggtcttctg
     1561 ttccccagag ccacctgcca agacctctac ccctgaagac ttcatccgaa tgaccaaggg
     1621 tattactatg gcaacagcca aagccgttgc tgctggcaat tcctgtcgac aggaagatgt
     1681 cattgccaca gccaatctga gccgacgggc tattgcggac atgcttcggg cttgcaagga
     1741 agcagctttc cacccagaag tggcgcctga tgtacggctc cgagccctgc actatggccg
     1801 ggagtgtgcc aatggttacc tggaactgct ggaccacgtg ctgctgaccc ttcagaagcc
     1861 aaacccagac ctgaagcagc agctaactgg acactcaaag cgggttgctg gctctgtgac
     1921 tgagctcatc caggctgctg aagccatgaa aggaacagag tgggtggacc cagaggaccc
     1981 tactgtcatt gctgagaatg agctcttggg agccgcagcc gccatcgagg ctgcagccaa
     2041 gaagctggag cagctgaagc ccagggccaa acccaaggag gcggatgagt ccttgaactt
     2101 tgaggaacaa gggagtcctg gcggtggatc cggaggctct gggagctctg ccagcggtgg
     2161 cagcacctcc gatctgcctg atgaccacta cctgagcaca cagaccatcc tctccaagga
     2221 ccttaactaa gaattcagat ccgcccctct ccctcccccc cccctaacgt tactggccga
     2281 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     2341 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     2401 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     2461 cctctggtag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     2521 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     2581 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     2641 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     2701 ggatctgatc tggggcctcg gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa
     2761 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     2821 ggccacaacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     2881 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     2941 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     3001 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     3061 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     3121 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     3181 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     3241 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     3301 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     3361 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     3421 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     3481 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     3541 aaccggtgct cgaggtaccg atgaatagct aaggtcgagg ccgcaggtaa gtatcaaggt
     3601 tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt
     3661 gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg
     3721 tgtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3781 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3841 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3901 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3961 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4021 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4081 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4141 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4201 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4261 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4321 aattcgagct cggtacgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4381 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4441 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4501 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcccagcttt tgttcccttt
     4561 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4621 gttatccgct aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     4681 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     4741 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     4801 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     4861 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     4921 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     4981 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     5041 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     5101 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcg
     5161 gccggccgcc ggcgtcgacg cgcgcggccg cggcgcgccg ttgctggcgt ttttccatag
     5221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5641 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6961 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7021 gccctttcgt ctacgcgcgc gccggcggcg cgccgtcgac ggccggccgc ggccgc
//
