LOCUS       sequence-249236-        6180 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 6180)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6180)
  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-355 C-Tag Venus173/155[2] L10 {Esp3I(2x)...
FEATURES             Location/Qualifiers
     source          1..6180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     733..757
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    join(1348..1922,1923..1925,1926..1934)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1515..1532)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1742..1761
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1935..2645
                     /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(2080..2098)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2348..2367
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2691..2823
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2832..3420
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2885..2905)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3303..3314)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3447..3464)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3453..3564
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3660..3680)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3660..3678)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3699..3715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3699..3715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3712..3734)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3817..3836)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3882..3937
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3936..3955)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4316..4904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4396..4415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5075..5866,5867..5935))
                     /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     5698..5717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5936..6040)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6108..6126
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcctgcctcc aggcctcaat attggccatt agccatatta ttcattggtt atatagcata
       61 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
      121 tattggctca tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt
      781 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag
      841 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc
     1021 cacccgagac gccgaggcac gccgtctccg ggagctctgc cagcggtggc agcaccagtg
     1081 ataagcagaa gaatgggatt aaggcgaact tcaagatccg acacaacatc gaggatggtg
     1141 gggtccagct tgccgaccat tatcagcaga acacacccat tggggatgga ccagtcctgt
     1201 tgccggataa ccactacctc agttaccaga gcaaactgtc caaagatccc aatgagaaga
     1261 gggaccatat ggtactgctc gaattcgtga ctgcagcagg cattaccctt gggatggatg
     1321 aactttataa gtaagaattc agatccgccc ctctccctcc ccccccccta acgttactgg
     1381 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     1441 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     1501 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     1561 agttcctctg gtagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     1621 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     1681 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     1741 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     1801 tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa
     1861 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     1921 atatggccac aaccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     1981 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     2041 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     2101 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     2161 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     2221 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     2281 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     2341 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     2401 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2461 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2521 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     2581 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2641 agtaaaccgg tgctcgaggt accgatgaat agctaaggtc gaggccgcag gtaagtatca
     2701 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     2761 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     2821 caggtgtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2881 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2941 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3001 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3061 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3121 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3181 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     3241 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3301 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3361 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3421 ctggaattcg agctcggtac gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3481 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3541 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3601 tggggtgggg caggacagca agggggagga ttgggaagac aatagcccag cttttgttcc
     3661 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     3721 aattgttatc cgctaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     3781 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     3841 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     3901 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3961 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     4021 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     4081 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     4141 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     4201 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     4261 atcggccggc cgccggcgtc gacgcgcgcg gccgcggcgc gccgttgctg gcgtttttcc
     4321 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4741 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5041 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5101 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5161 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5221 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5281 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5341 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5401 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5461 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5521 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5581 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5641 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5701 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5761 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5821 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5881 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5941 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6001 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6061 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6121 cgaggccctt tcgtctacgc gcgcgccggc ggcgcgccgt cgacggccgg ccgcggccgc
//
