LOCUS       sequence-271322-        5707 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 5707)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5707)
  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-483 GFP[1] L9 Stuffer/Intein3 L10 GFP[2]...
FEATURES             Location/Qualifiers
     source          1..5707
                     /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(1649..2223,2224..2226,2227..2235)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1816..1833)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2043..2062
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2245..2949
                     /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(2387..2405)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2655..2674
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2956..3012
                     /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      4013..4042
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4064..4651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4552..4571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4663..4690)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4714..5505,5506..5574))
                     /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     5337..5356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5575..5677)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5678..5707,1..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 agtggaggcg accaactgac agaagagcag
     1021 attgcagagt tcaaagaagc cttctcatta ttcgacaagg atggggacgg caccatcacc
     1081 acaaaggaac ttggcaccgt tatgaggtcg cttggacaaa acccaacgga agcagaattg
     1141 caggatatga tcaatgaagt cgatgctgat ggcaatggaa cgatttactt tcctgaactt
     1201 cttactatgc tggctagaaa aatgaaggac acagacagcg aagaggaaat ccgagaagca
     1261 ttccgtgttt ttgacaagga tgggaacggc tacatcagcg ctgctgaatt acgtcacgtc
     1321 atgacaaacc tcggggagaa gttaacagat gaagaagttg atgaaatgat aagggaagca
     1381 gatatcgatg gtgatggcca agtaaactat gaagagtttg tacaaatgat gacagcaaag
     1441 ggggggaaga ggcgctggca gaaaacaggc catgctgtcc gaataatagg aagactgaag
     1501 aagatctcca gctccggggc actggcgtcg ggaggcagcg gtggtagtgg aggcgaaaag
     1561 agggaccaca tggtgctgct cgagtacgtc acagcagctg gcatcaccga tgccagctaa
     1621 tgcttagata agtgagaatt cagatccgcc cctctccctc ccccccccct aacgttactg
     1681 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     1741 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     1801 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     1861 cagttcctct ggtagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     1921 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     1981 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     2041 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     2101 gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa
     2161 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     2221 aatatggcca caaccatggg tgcagtgagc aagggcgagg aggataacat ggccatcatc
     2281 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     2341 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     2401 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     2461 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     2521 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     2581 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     2641 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     2701 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     2761 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     2821 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     2881 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     2941 ctgtacaaga gcggcgctac taacttcagc ctgctgaagc aggctggcga cgtggaggag
     3001 aaccctggac cttctggaag aaaaggcgaa gagctgttta caggggttgt gcctatcctc
     3061 attgagctcg atggagatgt caacggccac aagttctttg tgagagggga aggagaaggt
     3121 gacgctacta tcggcaaact gtctctgaag ttcatttgca ctaccgggaa gttgcccgta
     3181 ccatggccaa cactggtaac tacgctgaca tatggagtcc agtgtttcag ccgttatccg
     3241 gatcacatga agaggcatga cttctttaaa tccgcaatgc ctgagggata tgtccaagag
     3301 aggaccatct actttaagga cgacggtacc tacaaaaccc gagctgaagt gaagtttgag
     3361 ggtgacactc tggtgaatcg gattgagctc aaaggcatcg acttcaagga ggatggcaac
     3421 atcttgggcc ataagctgga gtacaacttc aacagtcaca aggtgtacat cacagccgac
     3481 aaacagaaca atgggattaa ggccaacttc accatacgcc acaatgtgga agatggctca
     3541 gtccaacttg cggaccatta ccagcagaat acgcctatag gggatggacc cgttcttctg
     3601 ccctaggttg ccttgagcat ctgacttctg gctaaattga tgatctatta aacaataaag
     3661 atgtccacat ggaagttttt tcctgtcata ctttgttaag aagggtgaga acagagtacc
     3721 tacattttga atggaaggat tggagctacg ggggtggggg tggggtggga ttagataaat
     3781 gcctgctctt tactgaaggc tctttactat tgctttatga taatgtttca tagttggatc
     3841 gaacacatat tgatcctcgt cacaaccaca ccagcacacc tcaaatccca caccactccc
     3901 acaattacca ttcactcaac aaactcacac atcccacgat aacgaattca agcttgatat
     3961 cattcaggac gagcctcaga ctccagcgta actggactgc aatcaactca ctggctcacc
     4021 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     4081 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4141 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     4201 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4261 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4321 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4381 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4441 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4501 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4561 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     4621 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     4681 aaggtgagcc aggtgattac atttgggccc tcattaccaa tgcttaatca gtgaggcacc
     4741 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4801 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc
     4861 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     4921 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4981 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5041 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5101 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5161 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5221 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     5281 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5341 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5401 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5461 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5521 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     5581 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     5641 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     5701 cttttga
//
