LOCUS       sequence-249406-        7106 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 7106)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7106)
  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-549    C-Tag Venus210[2] L20 Tln1(rod) m...
FEATURES             Location/Qualifiers
     source          1..7106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1018..1027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(2273..2847,2848..2850,2851..2859)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2440..2457)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2667..2686
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2860..3570
                     /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(3005..3023)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3273..3292
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          3616..3748
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3757..4345
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3810..3830)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4228..4239)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4372..4389)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4378..4489
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4585..4605)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4585..4603)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4624..4640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4624..4640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4637..4659)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4742..4761)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4807..4862
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4861..4880)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5241..5829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5321..5340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6000..6791,6792..6860))
                     /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     6623..6642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6861..6965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7033..7051
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctgcctcca ggcctcaata ttggccatta gccatattat tcattggtta tatagcataa
       61 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      121 attggctcat gtccaatatg accgccatgt tggcattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggcc
     1021 accatggctg tgtctggtat cattgctgac ctcgacacca ccatcatgtt cgctactgct
     1081 ggcacactta accgtgaggg tgctgaaact tttgctgacc accgggaggg tatcttaaag
     1141 acagcaaagg ttcttgtgga ggacaccaag gtcctagtgc agaatgcagc tgggagccag
     1201 gagaagttgg cacaagccgc ccagtcctcc gtggccacca ttacccgcct cgctgatgtg
     1261 gtcaagctcg gtgcagccag cctaggagcc gaagaccctg aaactcaggt ggtgctgatc
     1321 aatgcagtaa aggacgtagc caaggccctg ggtgacctca tcagcgctac gaaggctgca
     1381 gcgggcaaag ttggggatga ccctgcagtg tggcagctca agaactctgc caaggtgatg
     1441 gtgaccaatg tgacatcatt gctcaagaca gtgaaggctg tggaagatga ggccaccaaa
     1501 ggcacacggg ccctagaggc aaccacagag cacatacgtc aggaactggc ggtcttctgt
     1561 tccccagagc cacctgccaa gacctctacc cctgaagact tcatccgaat gaccaagggt
     1621 attactatgg caacagccaa agccgttgct gctggcaatt cctgtcgaca ggaagatgtc
     1681 attgccacag ccaatctgag ccgacgggct attgcggaca tgcttcgggc ttgcaaggaa
     1741 gcagctttcc acccagaagt ggcgcctgat gtacggctcc gagccctgca ctatggccgg
     1801 gagtgtgcca atggttacct ggaactgctg gaccacgtgc tgctgaccct tcagaagcca
     1861 aacccagacc tgaagcagca gctaactgga cactcaaagc gggttgctgg ctctgtgact
     1921 gagctcatcc aggctgctga agccatgaaa ggaacagagt gggtggaccc agaggaccct
     1981 actgtcattg ctgagaatga gctcttggga gccgcagccg ccatcgaggc tgcagccaag
     2041 aagctggagc agctgaagcc cagggccaaa cccaaggagg cggatgagtc cttgaacttt
     2101 gaggaacaag ggagtcctgg cggtggatcc ggaggctctg ggagctctgc cagcggtggc
     2161 agcaccagtg atcccaatga gaagagggac catatggtac tgctcgaatt cgtgactgca
     2221 gcaggcatta cccttgggat ggatgaactt tataagtaag aattcagatc cgcccctctc
     2281 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     2341 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     2401 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     2461 gtctgttgaa tgtcgtgaag gaagcagttc ctctggtagc ttcttgaaga caaacaacgt
     2521 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     2581 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     2641 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     2701 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     2761 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     2821 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     2881 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     2941 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3001 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     3061 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     3121 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3181 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     3241 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     3301 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3361 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3421 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     3481 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     3541 accggcggca tggacgagct gtacaagtaa accggtgctc gaggtaccga tgaatagcta
     3601 aggtcgaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
     3661 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
     3721 gacatccact ttgcctttct ctccacaggt gtcgacaatc aacctctgga ttacaaaatt
     3781 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3841 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3901 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3961 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4021 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4081 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4141 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     4201 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4261 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4321 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtacgatca gcctcgactg
     4381 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4441 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4501 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4561 aagacaatag cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     4621 atggtcatag ctgtttcctg tgtgaaattg ttatccgcta attcactcct caggtgcagg
     4681 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     4741 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     4801 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     4861 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     4921 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     4981 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     5041 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     5101 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     5161 tagctgtccc tcttctctta tggagatcgg ccggccgccg gcgtcgacgc gcgcggccgc
     5221 ggcgcgccgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5281 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5341 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5401 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5461 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5521 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5581 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5641 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5701 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5761 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5821 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5881 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5941 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6001 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6061 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6121 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6181 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6241 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6301 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6361 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6421 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6481 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6541 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6601 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6661 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6721 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6781 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6841 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6901 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6961 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7021 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tacgcgcgcg ccggcggcgc
     7081 gccgtcgacg gccggccgcg gccgcg
//
