LOCUS       sequence-271329-        5353 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 5353)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5353)
  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-489 GFP[1] L3 Stuffer/Intein1 L4 GFP[2] ...
FEATURES             Location/Qualifiers
     source          1..5353
                     /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(1295..1869,1870..1872,1873..1881)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1462..1479)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1689..1708
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1891..2595
                     /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(2033..2051)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2301..2320
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2602..2658
                     /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      3659..3688
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3710..4297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4198..4217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4309..4336)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4360..5151,5152..5220))
                     /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     4983..5002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5221..5323)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5324..5353,1..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 cagcgaagag gaactgagcg agtgcttcag aatcttcgac
     1021 ttcgacggca acggcttcat cgacagagag gagtttggcg acatcatcag actgaccggc
     1081 gagcagctga ccgacgagga cgtggacgag atcttcggcg actccgacac cgacaagaac
     1141 ggcagaatcg atttcgacga gttcctgaag atggtggaaa acgtgcaggc gtcgggaggc
     1201 gaaaagaggg accacatggt gctgctcgag tacgtcacag cagctggcat caccgatgcc
     1261 agctaatgct tagataagtg agaattcaga tccgcccctc tccctccccc ccccctaacg
     1321 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1381 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1441 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1501 aggaagcagt tcctctggta gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1561 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1621 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1681 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1741 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
     1801 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     1861 gatgataata tggccacaac catgggtgca gtgagcaagg gcgaggagga taacatggcc
     1921 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1981 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2041 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     2101 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2161 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2221 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2281 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2341 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     2401 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     2461 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     2521 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2581 gacgagctgt acaagagcgg cgctactaac ttcagcctgc tgaagcaggc tggcgacgtg
     2641 gaggagaacc ctggaccttc tggaagaaaa ggcgaagagc tgtttacagg ggttgtgcct
     2701 atcctcattg agctcgatgg agatgtcaac ggccacaagt tctttgtgag aggggaagga
     2761 gaaggtgacg ctactatcgg caaactgtct ctgaagttca tttgcactac cgggaagttg
     2821 cccgtaccat ggccaacact ggtaactacg ctgacatatg gagtccagtg tttcagccgt
     2881 tatccggatc acatgaagag gcatgacttc tttaaatccg caatgcctga gggatatgtc
     2941 caagagagga ccatctactt taaggacgac ggtacctaca aaacccgagc tgaagtgaag
     3001 tttgagggtg acactctggt gaatcggatt gagctcaaag gcatcgactt caaggaggat
     3061 ggcaacatct tgggccataa gctggagtac aacttcaaca gtcacaaggt gtacatcaca
     3121 gccgacaaac agaacaatgg gattaaggcc aacttcacca tacgccacaa tgtggaagat
     3181 ggctcagtcc aacttgcgga ccattaccag cagaatacgc ctatagggga tggacccgtt
     3241 cttctgccct aggttgcctt gagcatctga cttctggcta aattgatgat ctattaaaca
     3301 ataaagatgt ccacatggaa gttttttcct gtcatacttt gttaagaagg gtgagaacag
     3361 agtacctaca ttttgaatgg aaggattgga gctacggggg tgggggtggg gtgggattag
     3421 ataaatgcct gctctttact gaaggctctt tactattgct ttatgataat gtttcatagt
     3481 tggatcgaac acatattgat cctcgtcaca accacaccag cacacctcaa atcccacacc
     3541 actcccacaa ttaccattca ctcaacaaac tcacacatcc cacgataacg aattcaagct
     3601 tgatatcatt caggacgagc ctcagactcc agcgtaactg gactgcaatc aactcactgg
     3661 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     3721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3781 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     3841 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3901 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3961 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4021 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4081 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4141 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4201 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     4261 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     4321 cacccgaagg tgagccaggt gattacattt gggccctcat taccaatgct taatcagtga
     4381 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4441 gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4501 agaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga
     4561 gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4621 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4681 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4741 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4801 gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4861 taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac
     4921 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4981 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5041 ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5101 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5161 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5221 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     5281 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     5341 cggaggcttt tga
//
