LOCUS       sequence-249250-        6045 bp    DNA     circular SYN 15-APR-2025
DEFINITION  PPI (BiFC/TriFC) assay vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6045)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRDAX-369 C-Tag Venus210[2] L20 {Esp3I(2x)} mC...
FEATURES             Location/Qualifiers
     source          1..6045
                     /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"
     misc_feature    join(1212..1786,1787..1789,1790..1798)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1379..1396)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1606..1625
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1799..2509
                     /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(1944..1962)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2212..2231
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2555..2687
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2696..3284
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2749..2769)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3167..3178)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3311..3328)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3317..3428
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3524..3544)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3524..3542)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3563..3579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3563..3579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3576..3598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3681..3700)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3746..3801
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3800..3819)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4180..4768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4260..4279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4939..5730,5731..5799))
                     /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     5562..5581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5800..5904)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5972..5990
                     /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 acccgagacg ccgaggcacg ccgtctccgg gagtcctggc ggtggatccg gaggctctgg
     1081 gagctctgcc agcggtggca gcaccagtga tcccaatgag aagagggacc atatggtact
     1141 gctcgaattc gtgactgcag caggcattac ccttgggatg gatgaacttt ataagtaaga
     1201 attcagatcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     1261 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     1321 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     1381 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggtagct
     1441 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     1501 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     1561 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     1621 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     1681 gggcctcggt acacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     1741 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     1801 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     1861 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1921 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1981 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     2041 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     2101 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     2161 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     2221 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     2281 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     2341 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     2401 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     2461 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaaa ccggtgctcg
     2521 aggtaccgat gaatagctaa ggtcgaggcc gcaggtaagt atcaaggtta caagacaggt
     2581 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     2641 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tcgacaatca
     2701 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2761 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2821 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2881 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2941 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3001 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3061 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3121 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3181 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3241 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg
     3301 gtacgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3361 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3421 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3481 agcaaggggg aggattggga agacaatagc ccagcttttg ttccctttag tgagggttaa
     3541 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa
     3601 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     3661 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     3721 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     3781 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     3841 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     3901 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     3961 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     4021 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     4081 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatcggc cggccgccgg
     4141 cgtcgacgcg cgcggccgcg gcgcgccgtt gctggcgttt ttccataggc tccgcccccc
     4201 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4261 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4321 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4381 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4441 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4501 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4561 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4621 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4681 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4741 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4801 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4861 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4921 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4981 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5041 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5101 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5161 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5221 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5281 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5341 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5401 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5461 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5521 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5581 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5641 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5701 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5761 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5821 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5881 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5941 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6001 acgcgcgcgc cggcggcgcg ccgtcgacgg ccggccgcgg ccgcg
//
