LOCUS       sequence-271321-        5392 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 5392)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5392)
  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-482 GFP[1] L9 Stuffer/Intein2 L10 GFP[2]...
FEATURES             Location/Qualifiers
     source          1..5392
                     /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(1334..1908,1909..1911,1912..1920)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1501..1518)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1728..1747
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1930..2634
                     /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(2072..2090)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2340..2359
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2641..2697
                     /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      3698..3727
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3749..4336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4237..4256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4348..4375)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4399..5190,5191..5259))
                     /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     5022..5041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5260..5362)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5363..5392,1..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 cagcggtggt agtggaggca gcgaagaaga gctatcagag
     1021 tgctttcgca tttttgacaa ggatggaaat ggtttcatcg acagggagga atttggagac
     1081 atcatccgcc ttactggaga acagctcaca gatgaggatg tcgatgagat atttggagat
     1141 tcagacacag acaaaaatga cggaaggatt gactttgatg agttcctgaa gatggtggag
     1201 aatgtccagg cgtcgggagg cagcggtggt agtggaggcg aaaagaggga ccacatggtg
     1261 ctgctcgagt acgtcacagc agctggcatc accgatgcca gctaatgctt agataagtga
     1321 gaattcagat ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     1381 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     1441 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     1501 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggtag
     1561 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     1621 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     1681 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     1741 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     1801 tggggcctcg gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     1861 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     1921 atgggtgcag tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1981 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2041 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     2101 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     2161 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2221 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2281 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2341 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2401 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2461 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2521 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2581 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagagcggc
     2641 gctactaact tcagcctgct gaagcaggct ggcgacgtgg aggagaaccc tggaccttct
     2701 ggaagaaaag gcgaagagct gtttacaggg gttgtgccta tcctcattga gctcgatgga
     2761 gatgtcaacg gccacaagtt ctttgtgaga ggggaaggag aaggtgacgc tactatcggc
     2821 aaactgtctc tgaagttcat ttgcactacc gggaagttgc ccgtaccatg gccaacactg
     2881 gtaactacgc tgacatatgg agtccagtgt ttcagccgtt atccggatca catgaagagg
     2941 catgacttct ttaaatccgc aatgcctgag ggatatgtcc aagagaggac catctacttt
     3001 aaggacgacg gtacctacaa aacccgagct gaagtgaagt ttgagggtga cactctggtg
     3061 aatcggattg agctcaaagg catcgacttc aaggaggatg gcaacatctt gggccataag
     3121 ctggagtaca acttcaacag tcacaaggtg tacatcacag ccgacaaaca gaacaatggg
     3181 attaaggcca acttcaccat acgccacaat gtggaagatg gctcagtcca acttgcggac
     3241 cattaccagc agaatacgcc tataggggat ggacccgttc ttctgcccta ggttgccttg
     3301 agcatctgac ttctggctaa attgatgatc tattaaacaa taaagatgtc cacatggaag
     3361 ttttttcctg tcatactttg ttaagaaggg tgagaacaga gtacctacat tttgaatgga
     3421 aggattggag ctacgggggt gggggtgggg tgggattaga taaatgcctg ctctttactg
     3481 aaggctcttt actattgctt tatgataatg tttcatagtt ggatcgaaca catattgatc
     3541 ctcgtcacaa ccacaccagc acacctcaaa tcccacacca ctcccacaat taccattcac
     3601 tcaacaaact cacacatccc acgataacga attcaagctt gatatcattc aggacgagcc
     3661 tcagactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     3721 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3841 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3901 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3961 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4021 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4081 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4141 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4201 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4261 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     4321 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     4381 attacatttg ggccctcatt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4441 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga
     4501 gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc
     4561 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac
     4621 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4681 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4741 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4801 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt
     4861 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4921 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4981 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5041 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat
     5101 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5161 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5221 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga
     5281 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     5341 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt ga
//
