LOCUS       sequence-271330-        5356 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 5356)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5356)
  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-490 GFP[1] L3 Stuffer/Intein2 L4 GFP[2] ...
FEATURES             Location/Qualifiers
     source          1..5356
                     /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(1298..1872,1873..1875,1876..1884)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1465..1482)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1692..1711
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1894..2598
                     /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(2036..2054)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2304..2323
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2605..2661
                     /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      3662..3691
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3713..4300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4201..4220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4312..4339)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4363..5154,5155..5223))
                     /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     4986..5005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5224..5326)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5327..5356,1..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 cagcgaagaa gagctatcag agtgctttcg catttttgac
     1021 aaggatggaa atggtttcat cgacagggag gaatttggag acatcatccg ccttactgga
     1081 gaacagctca cagatgagga tgtcgatgag atatttggag attcagacac agacaaaaat
     1141 gacggaagga ttgactttga tgagttcctg aagatggtgg agaatgtcca ggcgtcggga
     1201 ggcgaaaaga gggaccacat ggtgctgctc gagtacgtca cagcagctgg catcaccgat
     1261 gccagctaat gcttagataa gtgagaattc agatccgccc ctctccctcc ccccccccta
     1321 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     1381 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     1441 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     1501 tgaaggaagc agttcctctg gtagcttctt gaagacaaac aacgtctgta gcgacccttt
     1561 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1621 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1681 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1741 taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt
     1801 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1861 cacgatgata atatggccac aaccatgggt gcagtgagca agggcgagga ggataacatg
     1921 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     1981 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2041 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     2101 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     2161 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     2221 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     2281 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     2341 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     2401 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     2461 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     2521 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     2581 atggacgagc tgtacaagag cggcgctact aacttcagcc tgctgaagca ggctggcgac
     2641 gtggaggaga accctggacc ttctggaaga aaaggcgaag agctgtttac aggggttgtg
     2701 cctatcctca ttgagctcga tggagatgtc aacggccaca agttctttgt gagaggggaa
     2761 ggagaaggtg acgctactat cggcaaactg tctctgaagt tcatttgcac taccgggaag
     2821 ttgcccgtac catggccaac actggtaact acgctgacat atggagtcca gtgtttcagc
     2881 cgttatccgg atcacatgaa gaggcatgac ttctttaaat ccgcaatgcc tgagggatat
     2941 gtccaagaga ggaccatcta ctttaaggac gacggtacct acaaaacccg agctgaagtg
     3001 aagtttgagg gtgacactct ggtgaatcgg attgagctca aaggcatcga cttcaaggag
     3061 gatggcaaca tcttgggcca taagctggag tacaacttca acagtcacaa ggtgtacatc
     3121 acagccgaca aacagaacaa tgggattaag gccaacttca ccatacgcca caatgtggaa
     3181 gatggctcag tccaacttgc ggaccattac cagcagaata cgcctatagg ggatggaccc
     3241 gttcttctgc cctaggttgc cttgagcatc tgacttctgg ctaaattgat gatctattaa
     3301 acaataaaga tgtccacatg gaagtttttt cctgtcatac tttgttaaga agggtgagaa
     3361 cagagtacct acattttgaa tggaaggatt ggagctacgg gggtgggggt ggggtgggat
     3421 tagataaatg cctgctcttt actgaaggct ctttactatt gctttatgat aatgtttcat
     3481 agttggatcg aacacatatt gatcctcgtc acaaccacac cagcacacct caaatcccac
     3541 accactccca caattaccat tcactcaaca aactcacaca tcccacgata acgaattcaa
     3601 gcttgatatc attcaggacg agcctcagac tccagcgtaa ctggactgca atcaactcac
     3661 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     3721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3781 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     3841 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4201 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     4261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     4321 gcccacccga aggtgagcca ggtgattaca tttgggccct cattaccaat gcttaatcag
     4381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4441 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     4501 gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     4561 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4801 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4861 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     4921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5041 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5221 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     5281 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     5341 ggtcggaggc ttttga
//
