LOCUS       sequence-298149-        7076 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 7076)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7076)
  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-526    N-Tag GFP[1] L20 Tln1(rod) mCherry
FEATURES             Location/Qualifiers
     source          1..7076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..78
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     regulatory      1461..1470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          1485..1617
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1732..1756)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1753..1956)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1782..1802)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1957..2336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2543..2561)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2629..2733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2734..2802,2803..3594)
                     /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(2952..2971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3765..4353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4254..4273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4714..4733
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4732..4787
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4833..4852
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     4935..4957
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4954..4970
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4954..4970
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4989..5009
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4991..5009
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    5105..5216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5205..5222
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    5249..5837
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5355..5366
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5764..5784
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     intron          5846..5978
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(6024..6734)
                     /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(6302..6321)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6571..6589
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    join(6735..6743,6744..6746,6747..7076,1..245)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6908..6927)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 ttgtttgtct tcaagaagct accagaggaa ctgcttcctt cacgacattc aacagacctt
       61 gcattccttt ggcgagaggg gaaagacccc taggaatgct cgtcaagaag acagggccag
      121 gtttccgggc cctcacattg ccaaaagacg gcaatatggt ggaaaataac atatagacaa
      181 acgcacaccg gccttattcc aagcggcttc ggccagtaac gttagggggg ggggagggag
      241 aggggcggat ctgaattctt attgttcctc aaagttcaag gactcatccg cctccttggg
      301 tttggccctg ggcttcagct gctccagctt cttggctgca gcctcgatgg cggctgcggc
      361 tcccaagagc tcattctcag caatgacagt agggtcctct gggtccaccc actctgttcc
      421 tttcatggct tcagcagcct ggatgagctc agtcacagag ccagcaaccc gctttgagtg
      481 tccagttagc tgctgcttca ggtctgggtt tggcttctga agggtcagca gcacgtggtc
      541 cagcagttcc aggtaaccat tggcacactc ccggccatag tgcagggctc ggagccgtac
      601 atcaggcgcc acttctgggt ggaaagctgc ttccttgcaa gcccgaagca tgtccgcaat
      661 agcccgtcgg ctcagattgg ctgtggcaat gacatcttcc tgtcgacagg aattgccagc
      721 agcaacggct ttggctgttg ccatagtaat acccttggtc attcggatga agtcttcagg
      781 ggtagaggtc ttggcaggtg gctctgggga acagaagacc gccagttcct gacgtatgtg
      841 ctctgtggtt gcctctaggg cccgtgtgcc tttggtggcc tcatcttcca cagccttcac
      901 tgtcttgagc aatgatgtca cattggtcac catcaccttg gcagagttct tgagctgcca
      961 cactgcaggg tcatccccaa ctttgcccgc tgcagccttc gtagcgctga tgaggtcacc
     1021 cagggccttg gctacgtcct ttactgcatt gatcagcacc acctgagttt cagggtcttc
     1081 ggctcctagg ctggctgcac cgagcttgac cacatcagcg aggcgggtaa tggtggccac
     1141 ggaggactgg gcggcttgtg ccaacttctc ctggctccca gctgcattct gcactaggac
     1201 cttggtgtcc tccacaagaa cctttgctgt ctttaagata ccctcccggt ggtcagcaaa
     1261 agtttcagca ccctcacggt taagtgtgcc agcagtagcg aacatgatgg tggtgtcgag
     1321 gtcagcaatg ataccagaca cagcggaggt gctgccaccg ctggcagagc tcccagagcc
     1381 tccggatcca ccgccaggac tcccgttaag gtccttggag aggatggtct gtgtgctcag
     1441 gtagtggtca tcaggcagat ccatggtggc ctgggagtgg acacctgtgg agagaaaggc
     1501 aaagtggatg tcagtaagac caataggtgc ctatcagaaa cgcaagagtc ttctctgtct
     1561 cgacaagccc agtttctatt ggtctcctta aacctgtctt gtaaccttga tacttacctg
     1621 cccagtgcct cacgaccaac ttctgcagct taagttcgag actgttgtgt cagaagcact
     1681 gactgcgtta gcaatttaac tgtgataaac taccgcaata aagcttctag tgatctgacg
     1741 gttcactaaa cgagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     1801 cgtcaacggg gcggggttat tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     1861 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     1921 tccacgccca ttggtgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     1981 gtagatgtac tgccaagtag gaaagtcccg taaggtcatg tactgggcat aatgccaggc
     2041 gggccattta ccgtcattga cgtcaatagg gggcggactt ggcatatgat acacttgatg
     2101 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     2161 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     2221 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     2281 atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgccaaca
     2341 tggcggtcat attggacatg agccaatata aatgtacata ttatgatata gatacaacgt
     2401 atgcaatggc caatagccaa tattgattta tgctatataa ccaatgaata atatggctaa
     2461 tggccaatat tgaggcctgg aggcaggcgc ggccgcggcc ggccgtcgac ggcgcgccgc
     2521 cggcgcgcgc gtagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     2581 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2641 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2701 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2761 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2821 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2881 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2941 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3001 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3061 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3121 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3181 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3241 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3301 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3361 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3421 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3481 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3541 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3601 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3661 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3721 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3781 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3841 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3901 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3961 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4021 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4081 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4141 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4201 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4261 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4321 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacggcgc gccgcggccg
     4381 cgcgcgtcga cgccggcggc cggccgatct ccataagaga agagggacag ctatgactgg
     4441 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     4501 tgttaaagaa aaaaataaca caaaacaaaa tataaaaaaa atctaacctc aagtcaaggc
     4561 ttttctatgg aataaggaat ggacagcagg gggctgtttc atatactgat gacctcttta
     4621 tagccaacct ttgttcatgg cagccagcat atgggcatat gttgccaaac tctaaaccaa
     4681 atactcattc tgatgtttta aatgatttgc cctcccatat gtccttccga gtgagagaca
     4741 caaaaaattc caacacacta ttgcaatgaa aataaatttc ctttattagc cagaagtcag
     4801 atgctcaagg ggcttcatga tgtccccata atttttggca gagggaaaaa gatctcagtg
     4861 gtatttgtga gccagggcat tggccacacc agccaccacc ttctgatagg cagcctgcac
     4921 ctgaggagtg aattagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4981 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct gggctattgt cttcccaatc
     5041 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     5101 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     5161 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     5221 ggctgatcgt accgagctcg aattccaggc ggggaggcgg cccaaaggga gatccgactc
     5281 gtctgagggc gaaggcgaag acgcggaaga ggccgcagag ccggcagcag gccgcgggaa
     5341 ggaaggtccg ctggattgag ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc
     5401 caggtggcaa cacaggcgag cagccatgga aaggacgtca gcttccccga caacaccacg
     5461 gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg
     5521 agttccgccg tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg
     5581 gtggcaatgc cccaaccagt gggggttgcg tcagcaaaca cagtgcacac cacgccacgt
     5641 tgcctgacaa cgggccacaa ctcctcataa agagacagca accaggattt atacaaggag
     5701 gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat
     5761 ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg
     5821 taatccagag gttgattgtc gacacctgtg gagagaaagg caaagtggat gtcagtaaga
     5881 ccaataggtg cctatcagaa acgcaagagt cttctctgtc tcgacaagcc cagtttctat
     5941 tggtctcctt aaacctgtct tgtaaccttg atacttacct gcggcctcga ccttagctat
     6001 tcatcggtac ctcgagcacc ggtttacttg tacagctcgt ccatgccgcc ggtggagtgg
     6061 cggccctcgg cgcgttcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg
     6121 tccaacttga tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag
     6181 gtggtcttga cctcagcgtc gtagtggccg ccgtccttca gcttcagcct ctgcttgatc
     6241 tcgcccttca gggcgccgtc ctcggggtac atccgctcgg aggaggcctc ccagcccatg
     6301 gtcttcttct gcattacggg gccgtcggag gggaagttgg tgccgcgcag cttcaccttg
     6361 tagatgaact cgccgtcctg cagggaggag tcctgggtca cggtcaccac gccgccgtcc
     6421 tcgaagttca tcacgcgctc ccacttgaag ccctcgggga aggacagctt caagtagtcg
     6481 gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt acatgaactg aggggacagg
     6541 atgtcccagg cgaagggcag ggggccaccc ttggtcacct tcagcttggc ggtctgggtg
     6601 ccctcgtagg ggcggccctc gccctcgccc tcgatctcga actcgtggcc gttcacggag
     6661 ccctccatgt gcaccttgaa gcgcatgaac tccttgatga tggccatgtt atcctcctcg
     6721 cccttgctca ccatggttgt ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc
     6781 cccgtggttc ggggggccta gacgtttttt taacctcgac taaacacatg taaagcatgt
     6841 gtaccgaggc cccagatcag atcccataca atggggtacc ttctgggcat ccttcagccc
     6901 cttgttgaat acgcttgagg agagccattt gactctttcc acaactatcc aactcacaac
     6961 gtggcactgg ggttgtgccg cctttgcagg tgtatcttat acacgtggct tttggccgca
     7021 gaggcacctg tcgccaggtg gggggttccg ctgcctgcaa agggtcgcta cagacg
//
