LOCUS       sequence-271326-        5374 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 5374)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5374)
  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-486 GFP[1] L6 Stuffer/Intein2 L7 GFP[2] ...
FEATURES             Location/Qualifiers
     source          1..5374
                     /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(1316..1890,1891..1893,1894..1902)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1483..1500)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1710..1729
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1912..2616
                     /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(2054..2072)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2322..2341
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2623..2679
                     /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      3680..3709
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3731..4318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4219..4238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4330..4357)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4381..5172,5173..5241))
                     /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     5004..5023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5242..5344)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5345..5374,1..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 acagcggtgg tagtggaggc agcgaagaag agctatcaga gtgctttcgc
     1021 atttttgaca aggatggaaa tggtttcatc gacagggagg aatttggaga catcatccgc
     1081 cttactggag aacagctcac agatgaggat gtcgatgaga tatttggaga ttcagacaca
     1141 gacaaaaatg acggaaggat tgactttgat gagttcctga agatggtgga gaatgtccag
     1201 gcgtcgggag gcagcggtgg tgaaaagagg gaccacatgg tgctgctcga gtacgtcaca
     1261 gcagctggca tcaccgatgc cagctaatgc ttagataagt gagaattcag atccgcccct
     1321 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     1381 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     1441 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     1501 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctggt agcttcttga agacaaacaa
     1561 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     1621 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     1681 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     1741 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat
     1801 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     1861 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatgggtgc agtgagcaag
     1921 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     1981 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2041 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     2101 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     2161 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2221 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     2281 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     2341 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     2401 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     2461 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     2521 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     2581 cgccactcca ccggcggcat ggacgagctg tacaagagcg gcgctactaa cttcagcctg
     2641 ctgaagcagg ctggcgacgt ggaggagaac cctggacctt ctggaagaaa aggcgaagag
     2701 ctgtttacag gggttgtgcc tatcctcatt gagctcgatg gagatgtcaa cggccacaag
     2761 ttctttgtga gaggggaagg agaaggtgac gctactatcg gcaaactgtc tctgaagttc
     2821 atttgcacta ccgggaagtt gcccgtacca tggccaacac tggtaactac gctgacatat
     2881 ggagtccagt gtttcagccg ttatccggat cacatgaaga ggcatgactt ctttaaatcc
     2941 gcaatgcctg agggatatgt ccaagagagg accatctact ttaaggacga cggtacctac
     3001 aaaacccgag ctgaagtgaa gtttgagggt gacactctgg tgaatcggat tgagctcaaa
     3061 ggcatcgact tcaaggagga tggcaacatc ttgggccata agctggagta caacttcaac
     3121 agtcacaagg tgtacatcac agccgacaaa cagaacaatg ggattaaggc caacttcacc
     3181 atacgccaca atgtggaaga tggctcagtc caacttgcgg accattacca gcagaatacg
     3241 cctatagggg atggacccgt tcttctgccc taggttgcct tgagcatctg acttctggct
     3301 aaattgatga tctattaaac aataaagatg tccacatgga agttttttcc tgtcatactt
     3361 tgttaagaag ggtgagaaca gagtacctac attttgaatg gaaggattgg agctacgggg
     3421 gtgggggtgg ggtgggatta gataaatgcc tgctctttac tgaaggctct ttactattgc
     3481 tttatgataa tgtttcatag ttggatcgaa cacatattga tcctcgtcac aaccacacca
     3541 gcacacctca aatcccacac cactcccaca attaccattc actcaacaaa ctcacacatc
     3601 ccacgataac gaattcaagc ttgatatcat tcaggacgag cctcagactc cagcgtaact
     3661 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     3721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     3841 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3901 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3961 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4021 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4081 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4141 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4201 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4261 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4321 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     4381 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4441 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     4501 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     4561 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     4621 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4681 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4741 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4801 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     4861 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     4921 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4981 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5041 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     5101 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5161 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5221 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     5281 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     5341 atcaagtcaa aagcctccgg tcggaggctt ttga
//
