LOCUS       sequence-271331-        5671 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 5671)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5671)
  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-491 GFP[1] L3 Stuffer/Intein3 L4 GFP[2] ...
FEATURES             Location/Qualifiers
     source          1..5671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        185..564
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        566..841
                     /label=chicken beta-actin promoter
     regulatory      907..916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(1613..2187,2188..2190,2191..2199)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1780..1797)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2007..2026
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2209..2913
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2351..2369)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2619..2638
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2920..2976
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     terminator      3977..4006
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4028..4615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4516..4535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4627..4654)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4678..5469,5470..5538))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5301..5320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5539..5641)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5642..5671,1..2))
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctttctgcta tggaggtcag gtatgattta aatggtcagt attgagcgat atctagagaa
       61 ttcgtatagg tcttgtattc agtctcgcac tttagacatg gtcttctgct gcccgtggtc
      121 ctctattcgt cggatgttct gtcgattctt cctgtctatc tggactggat ctgatatcat
      181 cgtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      241 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      301 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      361 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      421 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      481 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      541 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      601 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      661 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      721 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      781 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      841 gggagtcgct gcgttgggac ttagccttta gtgaaccgtc agaattaatt cagatcgatc
      901 tacagcgcca ccatggatct gcctgatgac cactacctga gcacacagac catcctctcc
      961 aaggacctta acagtggagg cgaccaactg acagaagagc agattgcaga gttcaaagaa
     1021 gccttctcat tattcgacaa ggatggggac ggcaccatca ccacaaagga acttggcacc
     1081 gttatgaggt cgcttggaca aaacccaacg gaagcagaat tgcaggatat gatcaatgaa
     1141 gtcgatgctg atggcaatgg aacgatttac tttcctgaac ttcttactat gctggctaga
     1201 aaaatgaagg acacagacag cgaagaggaa atccgagaag cattccgtgt ttttgacaag
     1261 gatgggaacg gctacatcag cgctgctgaa ttacgtcacg tcatgacaaa cctcggggag
     1321 aagttaacag atgaagaagt tgatgaaatg ataagggaag cagatatcga tggtgatggc
     1381 caagtaaact atgaagagtt tgtacaaatg atgacagcaa agggggggaa gaggcgctgg
     1441 cagaaaacag gccatgctgt ccgaataata ggaagactga agaagatctc cagctccggg
     1501 gcactggcgt cgggaggcga aaagagggac cacatggtgc tgctcgagta cgtcacagca
     1561 gctggcatca ccgatgccag ctaatgctta gataagtgag aattcagatc cgcccctctc
     1621 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     1681 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     1741 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     1801 gtctgttgaa tgtcgtgaag gaagcagttc ctctggtagc ttcttgaaga caaacaacgt
     1861 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     1921 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     1981 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     2041 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     2101 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     2161 ttttcctttg aaaaacacga tgataatatg gccacaacca tgggtgcagt gagcaagggc
     2221 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2281 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2341 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2401 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2461 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2521 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2581 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2641 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2701 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2761 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2821 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2881 cactccaccg gcggcatgga cgagctgtac aagagcggcg ctactaactt cagcctgctg
     2941 aagcaggctg gcgacgtgga ggagaaccct ggaccttctg gaagaaaagg cgaagagctg
     3001 tttacagggg ttgtgcctat cctcattgag ctcgatggag atgtcaacgg ccacaagttc
     3061 tttgtgagag gggaaggaga aggtgacgct actatcggca aactgtctct gaagttcatt
     3121 tgcactaccg ggaagttgcc cgtaccatgg ccaacactgg taactacgct gacatatgga
     3181 gtccagtgtt tcagccgtta tccggatcac atgaagaggc atgacttctt taaatccgca
     3241 atgcctgagg gatatgtcca agagaggacc atctacttta aggacgacgg tacctacaaa
     3301 acccgagctg aagtgaagtt tgagggtgac actctggtga atcggattga gctcaaaggc
     3361 atcgacttca aggaggatgg caacatcttg ggccataagc tggagtacaa cttcaacagt
     3421 cacaaggtgt acatcacagc cgacaaacag aacaatggga ttaaggccaa cttcaccata
     3481 cgccacaatg tggaagatgg ctcagtccaa cttgcggacc attaccagca gaatacgcct
     3541 ataggggatg gacccgttct tctgccctag gttgccttga gcatctgact tctggctaaa
     3601 ttgatgatct attaaacaat aaagatgtcc acatggaagt tttttcctgt catactttgt
     3661 taagaagggt gagaacagag tacctacatt ttgaatggaa ggattggagc tacgggggtg
     3721 ggggtggggt gggattagat aaatgcctgc tctttactga aggctcttta ctattgcttt
     3781 atgataatgt ttcatagttg gatcgaacac atattgatcc tcgtcacaac cacaccagca
     3841 cacctcaaat cccacaccac tcccacaatt accattcact caacaaactc acacatccca
     3901 cgataacgaa ttcaagcttg atatcattca ggacgagcct cagactccag cgtaactgga
     3961 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     4021 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4081 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     4141 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4561 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4621 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatta
     4681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4741 tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag
     4801 tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca
     4861 gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc
     4921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4981 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5101 tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat
     5161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt
     5221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5281 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5341 cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag
     5401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5521 gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     5581 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     5641 aagtcaaaag cctccggtcg gaggcttttg a
//
