LOCUS       sequence-271327-        5689 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 5689)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5689)
  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-487 GFP[1] L6 Stuffer/Intein3 L7 GFP[2] ...
FEATURES             Location/Qualifiers
     source          1..5689
                     /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(1631..2205,2206..2208,2209..2217)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1798..1815)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2025..2044
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2227..2931
                     /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(2369..2387)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2637..2656
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2938..2994
                     /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      3995..4024
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4046..4633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4534..4553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4645..4672)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4696..5487,5488..5556))
                     /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     5319..5338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5557..5659)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5660..5689,1..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 gaccaactga cagaagagca gattgcagag
     1021 ttcaaagaag ccttctcatt attcgacaag gatggggacg gcaccatcac cacaaaggaa
     1081 cttggcaccg ttatgaggtc gcttggacaa aacccaacgg aagcagaatt gcaggatatg
     1141 atcaatgaag tcgatgctga tggcaatgga acgatttact ttcctgaact tcttactatg
     1201 ctggctagaa aaatgaagga cacagacagc gaagaggaaa tccgagaagc attccgtgtt
     1261 tttgacaagg atgggaacgg ctacatcagc gctgctgaat tacgtcacgt catgacaaac
     1321 ctcggggaga agttaacaga tgaagaagtt gatgaaatga taagggaagc agatatcgat
     1381 ggtgatggcc aagtaaacta tgaagagttt gtacaaatga tgacagcaaa gggggggaag
     1441 aggcgctggc agaaaacagg ccatgctgtc cgaataatag gaagactgaa gaagatctcc
     1501 agctccgggg cactggcgtc gggaggcagc ggtggtgaaa agagggacca catggtgctg
     1561 ctcgagtacg tcacagcagc tggcatcacc gatgccagct aatgcttaga taagtgagaa
     1621 ttcagatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     1681 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     1741 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     1801 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggtagctt
     1861 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     1921 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     1981 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2041 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2101 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2161 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     2221 ggtgcagtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2281 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2341 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2401 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2461 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2521 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2581 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2641 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2701 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2761 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2821 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2881 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gagcggcgct
     2941 actaacttca gcctgctgaa gcaggctggc gacgtggagg agaaccctgg accttctgga
     3001 agaaaaggcg aagagctgtt tacaggggtt gtgcctatcc tcattgagct cgatggagat
     3061 gtcaacggcc acaagttctt tgtgagaggg gaaggagaag gtgacgctac tatcggcaaa
     3121 ctgtctctga agttcatttg cactaccggg aagttgcccg taccatggcc aacactggta
     3181 actacgctga catatggagt ccagtgtttc agccgttatc cggatcacat gaagaggcat
     3241 gacttcttta aatccgcaat gcctgaggga tatgtccaag agaggaccat ctactttaag
     3301 gacgacggta cctacaaaac ccgagctgaa gtgaagtttg agggtgacac tctggtgaat
     3361 cggattgagc tcaaaggcat cgacttcaag gaggatggca acatcttggg ccataagctg
     3421 gagtacaact tcaacagtca caaggtgtac atcacagccg acaaacagaa caatgggatt
     3481 aaggccaact tcaccatacg ccacaatgtg gaagatggct cagtccaact tgcggaccat
     3541 taccagcaga atacgcctat aggggatgga cccgttcttc tgccctaggt tgccttgagc
     3601 atctgacttc tggctaaatt gatgatctat taaacaataa agatgtccac atggaagttt
     3661 tttcctgtca tactttgtta agaagggtga gaacagagta cctacatttt gaatggaagg
     3721 attggagcta cgggggtggg ggtggggtgg gattagataa atgcctgctc tttactgaag
     3781 gctctttact attgctttat gataatgttt catagttgga tcgaacacat attgatcctc
     3841 gtcacaacca caccagcaca cctcaaatcc cacaccactc ccacaattac cattcactca
     3901 acaaactcac acatcccacg ataacgaatt caagcttgat atcattcagg acgagcctca
     3961 gactccagcg taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc
     4021 ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4081 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4141 ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4201 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4261 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4321 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4381 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4441 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4501 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4561 tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg
     4621 gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt
     4681 acatttgggc cctcattacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg
     4801 cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga
     4861 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt
     4921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc
     5161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5221 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt
     5401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5521 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa
     5581 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     5641 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttga
//
