LOCUS       sequence-298151-        7112 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 7112)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7112)
  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-527    C-Tag GFP[1] L32 Tln1(rod) mCherry
FEATURES             Location/Qualifiers
     source          1..7112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..87
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     regulatory      1506..1515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          1530..1662
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1777..1801)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1798..2001)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1827..1847)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2002..2381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2588..2606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2674..2778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2779..2847,2848..3639)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2997..3016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3810..4398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4299..4318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4759..4778
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4777..4832
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4878..4897
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     4980..5002
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4999..5015
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4999..5015
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5034..5054
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5036..5054
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    5150..5261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5250..5267
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    5294..5882
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5400..5411
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5809..5829
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     intron          5891..6023
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(6069..6779)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6347..6366)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6616..6634
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    join(6780..6788,6789..6791,6792..7112,1..254)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6953..6972)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 ctacagacgt tgtttgtctt caagaagcta ccagaggaac tgcttccttc acgacattca
       61 acagaccttg cattcctttg gcgagagggg aaagacccct aggaatgctc gtcaagaaga
      121 cagggccagg tttccgggcc ctcacattgc caaaagacgg caatatggtg gaaaataaca
      181 tatagacaaa cgcacaccgg ccttattcca agcggcttcg gccagtaacg ttaggggggg
      241 gggagggaga ggggcggatc tgaattctta gttaaggtcc ttggagagga tggtctgtgt
      301 gctcaggtag tggtcatcag gcagatcact cccaccaccg gctgacccgc ctccagcgct
      361 cccctctccg ccagagccag gccctccgga tccgccgcct tcgctccctc cgccacccac
      421 gtcttgttcc tcaaagttca aggactcatc cgcctccttg ggtttggccc tgggcttcag
      481 ctgctccagc ttcttggctg cagcctcgat ggcggctgcg gctcccaaga gctcattctc
      541 agcaatgaca gtagggtcct ctgggtccac ccactctgtt cctttcatgg cttcagcagc
      601 ctggatgagc tcagtcacag agccagcaac ccgctttgag tgtccagtta gctgctgctt
      661 caggtctggg tttggcttct gaagggtcag cagcacgtgg tccagcagtt ccaggtaacc
      721 attggcacac tcccggccat agtgcagggc tcggagccgt acatcaggcg ccacttctgg
      781 gtggaaagct gcttccttgc aagcccgaag catgtccgca atagcccgtc ggctcagatt
      841 ggctgtggca atgacatctt cctgtcgaca ggaattgcca gcagcaacgg ctttggctgt
      901 tgccatagta atacccttgg tcattcggat gaagtcttca ggggtagagg tcttggcagg
      961 tggctctggg gaacagaaga ccgccagttc ctgacgtatg tgctctgtgg ttgcctctag
     1021 ggcccgtgtg cctttggtgg cctcatcttc cacagccttc actgtcttga gcaatgatgt
     1081 cacattggtc accatcacct tggcagagtt cttgagctgc cacactgcag ggtcatcccc
     1141 aactttgccc gctgcagcct tcgtagcgct gatgaggtca cccagggcct tggctacgtc
     1201 ctttactgca ttgatcagca ccacctgagt ttcagggtct tcggctccta ggctggctgc
     1261 accgagcttg accacatcag cgaggcgggt aatggtggcc acggaggact gggcggcttg
     1321 tgccaacttc tcctggctcc cagctgcatt ctgcactagg accttggtgt cctccacaag
     1381 aacctttgct gtctttaaga taccctcccg gtggtcagca aaagtttcag caccctcacg
     1441 gttaagtgtg ccagcagtag cgaacatgat ggtggtgtcg aggtcagcaa tgataccaga
     1501 cacagccatg gtggcctggg agtggacacc tgtggagaga aaggcaaagt ggatgtcagt
     1561 aagaccaata ggtgcctatc agaaacgcaa gagtcttctc tgtctcgaca agcccagttt
     1621 ctattggtct ccttaaacct gtcttgtaac cttgatactt acctgcccag tgcctcacga
     1681 ccaacttctg cagcttaagt tcgagactgt tgtgtcagaa gcactgactg cgttagcaat
     1741 ttaactgtga taaactaccg caataaagct tctagtgatc tgacggttca ctaaacgagc
     1801 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca acggggcggg
     1861 gttattacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     1921 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattggt
     1981 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     2041 agtaggaaag tcccgtaagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     2101 attgacgtca atagggggcg gacttggcat atgatacact tgatgtactg ccaagtgggc
     2161 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     2221 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     2281 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     2341 cgtaattgat tactattaat aactagtcaa taatcaatgc caacatggcg gtcatattgg
     2401 acatgagcca atataaatgt acatattatg atatagatac aacgtatgca atggccaata
     2461 gccaatattg atttatgcta tataaccaat gaataatatg gctaatggcc aatattgagg
     2521 cctggaggca ggcgcggccg cggccggccg tcgacggcgc gccgccggcg cgcgcgtaga
     2581 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2641 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2701 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2761 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2821 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2881 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2941 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3001 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3061 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3121 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3181 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3241 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3301 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3361 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3421 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3481 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3541 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3601 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3661 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3721 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3781 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4381 gggcggagcc tatggaaaaa cgccagcaac ggcgcgccgc ggccgcgcgc gtcgacgccg
     4441 gcggccggcc gatctccata agagaagagg gacagctatg actgggagta gtcaggagag
     4501 gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa
     4561 taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc tatggaataa
     4621 ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc aacctttgtt
     4681 catggcagcc agcatatggg catatgttgc caaactctaa accaaatact cattctgatg
     4741 ttttaaatga tttgccctcc catatgtcct tccgagtgag agacacaaaa aattccaaca
     4801 cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct caaggggctt
     4861 catgatgtcc ccataatttt tggcagaggg aaaaagatct cagtggtatt tgtgagccag
     4921 ggcattggcc acaccagcca ccaccttctg ataggcagcc tgcacctgag gagtgaatta
     4981 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     5041 accctcacta aagggaacaa aagctgggct attgtcttcc caatcctccc ccttgctgtc
     5101 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     5161 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     5221 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcgtaccga
     5281 gctcgaattc caggcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg
     5341 cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga
     5401 ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag
     5461 gcgagcagcc atggaaagga cgtcagcttc cccgacaaca ccacggaatt gtcagtgccc
     5521 aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca
     5581 atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa
     5641 ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc
     5701 cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc
     5761 atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa
     5821 ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga
     5881 ttgtcgacac ctgtggagag aaaggcaaag tggatgtcag taagaccaat aggtgcctat
     5941 cagaaacgca agagtcttct ctgtctcgac aagcccagtt tctattggtc tccttaaacc
     6001 tgtcttgtaa ccttgatact tacctgcggc ctcgacctta gctattcatc ggtacctcga
     6061 gcaccggttt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
     6121 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg
     6181 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     6241 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     6301 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
     6361 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     6421 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     6481 cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     6541 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     6601 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     6661 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     6721 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg
     6781 gttgtggcca tattatcatc gtgtttttca aaggaaaacc acgtccccgt ggttcggggg
     6841 gcctagacgt ttttttaacc tcgactaaac acatgtaaag catgtgtacc gaggccccag
     6901 atcagatccc atacaatggg gtaccttctg ggcatccttc agccccttgt tgaatacgct
     6961 tgaggagagc catttgactc tttccacaac tatccaactc acaacgtggc actggggttg
     7021 tgccgccttt gcaggtgtat cttatacacg tggcttttgg ccgcagaggc acctgtcgcc
     7081 aggtgggggg ttccgctgcc tgcaaagggt cg
//
