LOCUS       sequence-249421-        8420 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 8420)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8420)
  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-516    N-Tag GFP[1] L32 Itgb1 mCherry
FEATURES             Location/Qualifiers
     source          1..8420
                     /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"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(3588..4162,4163..4165,4166..4174)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3755..3772)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3982..4001
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4175..4885
                     /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(4320..4338)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4588..4607
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4931..5063
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5072..5660
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5125..5145)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5543..5554)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5687..5704)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5693..5804
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5900..5920)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5900..5918)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5939..5955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5939..5955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5952..5974)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6057..6076)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6122..6177
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6176..6195)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6556..7144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6636..6655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7315..8106,8107..8175))
                     /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     7938..7957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8176..8280)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8348..8366
                     /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 caccatggat ctgcctgatg accactacct gagcacacag accatcctct ccaaggacct
     1081 taacgacgtg ggtggcggag ggagcgaagg cggcggatcc ggagggcctg gctctggcgg
     1141 agaggggagc gctggaggcg ggtcagccgg tggtgggagt aatttgcaac tggtttcctg
     1201 gattggattg atcagtttga tttgttctgt atttggccaa acagataaaa atagatgttt
     1261 aaaagcaaat gccaaatctt gcggagaatg tatacaagca gggccaaatt gtgggtggtg
     1321 tacaaatacg acatttttgc aagaaggaat gcctacttct gcacgatgtg atgatttaga
     1381 agctttgaaa aagaagggtt gccagccaag tgacatagag aatcccagag gctctcaaac
     1441 tataaagaaa aataaaaatg tcaccaatcg cagcaaaggg atggcagaga agctccggcc
     1501 agaagacatt actcagatcc aaccacaaca gctgcttcta aaattgagat caggagaacc
     1561 acagaagttt acattaaaat tcaagagggc tgaagattac cctattgatc tctactacct
     1621 tatggatctc tcctactcta tgaaagatga tctggagaat gtgaaaagtc ttggaacgga
     1681 tttgatgaat gaaatgagga ggattacttc agacttccgc attggctttg gctcatttgt
     1741 ggagaaaact gtgatgccgt atattagcac aaccccagca aagctaagaa atccttgtac
     1801 aagtgaacaa aactgcacca gcccatttag ctacaaaaat gtgcttagtc ttactgacag
     1861 aggagagttt ttcaatgaac ttgttggtca gcaacgcata tctggaaact tggattctcc
     1921 agaaggtggc tttgatgcaa tcatgcaggt tgcggtttgt ggatcgctga ttggctggag
     1981 gaatgtaaca cgactgctgg tgttttccac ggatgctggg tttcactttg ctggagatgg
     2041 gaaacttggt ggtattgttt tacccaatga tggacaatgt cacctggaaa ataatgtata
     2101 tacaatgagc cattactatg attatccttc aattgctcac cttgttcaga aactaagtga
     2161 aaataatatt cagacgattt ttgcagttac tgaagagttc caacctgttt acaaggaatt
     2221 gaagaatttg attcctaagt cagcagtggg cacactgtct ggaaactcta gtaatgtgat
     2281 ccagctaatc atcgatgcct acaactctct ttcttcagaa gtcattctgg aaaatagcaa
     2341 attgccagac ggagtaacaa taaattacaa atcctattgc aagaatgggg tgaatgggac
     2401 aggagaaaat ggacgaaagt gttccaacat ttctattgga gatgaggttc aatttgaaat
     2461 tagcataact gctaataaat gtccaaataa ggagtctgaa accattaaaa ttaaacctct
     2521 gggcttcact gaagaagtag aggtcgttct tcagttcatc tgtaagtgca attgtcaaag
     2581 ccatggcatc ccagccagtc ccaagtgcca tgagggaaat gggacatttg agtgtggagc
     2641 ctgcaggtgc aatgaggggc gtgttgggag gcactgtgaa tgtagcacag atgaagtgaa
     2701 cagtgaagac atggacgctt actgcaggaa agagaacagt tcggaaatct gcagtaataa
     2761 tggagaatgt gtctgtggac agtgtgtgtg taggaagaga gataatacaa atgaaattta
     2821 ctctggaaaa ttctgcgagt gtgataactt caactgtgat aggtctaatg gcttaatttg
     2881 tggaggcaat ggcgtgtgca ggtgtcgtgt ttgtgaatgc tatcccaatt acactggcag
     2941 tgcatgtgac tgttctttgg acactggtcc atgtctagcg tcaaatggtc agatctgcaa
     3001 tggccggggt atttgtgaat gtggtgcttg taagtgcaca gatcccaagt ttcaagggcc
     3061 aacttgtgag acatgtcaga cctgccttgg cgtctgtgca gagcataaag aatgtgttca
     3121 gtgcagagcc ttcaataaag gagaaaagaa agacacgtgt gcacaggagt gctcccactt
     3181 caatctcacc aaagtagaaa gcagggagaa gttgccccag ccggtgcagg tcgatcctgt
     3241 gacccattgc aaggagaagg acattgatga ctgctggttc tatttcacct attcagtgaa
     3301 tggcaacaat gaagctatcg tgcatgttgt ggagactcca gactgtccta ctggtcccga
     3361 catcatccca attgtagcag gcgtggttgc tggaattgtt cttattggcc ttgccttgct
     3421 gctgatttgg aaacttttaa tgataattca tgacagaagg gaatttgcta aatttgaaaa
     3481 ggagaaaatg aatgccaagt gggacacggg tgaaaatcct atttacaaga gtgccgtgac
     3541 aactgtggtc aatccgaagt atgagggaaa ataagaattc agatccgccc ctctccctcc
     3601 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3661 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3721 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3781 ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt gaagacaaac aacgtctgta
     3841 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3901 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3961 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     4021 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     4081 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4141 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggaggataa
     4201 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     4261 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     4321 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     4381 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     4441 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     4501 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     4561 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4621 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     4681 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     4741 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     4801 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     4861 cggcatggac gagctgtaca agtaaaccgg tgctcgaggt accgatgaat agctaaggtc
     4921 gaggccgcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     4981 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
     5041 ccactttgcc tttctctcca caggtgtcga caatcaacct ctggattaca aaatttgtga
     5101 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5161 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5221 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5281 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5341 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5401 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5461 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5521 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5581 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5641 cctttgggcc gcctccccgc ctggaattcg agctcggtac gatcagcctc gactgtgcct
     5701 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     5761 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     5821 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     5881 aatagcccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5941 catagctgtt tcctgtgtga aattgttatc cgctaattca ctcctcaggt gcaggctgcc
     6001 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     6061 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     6121 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     6181 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     6241 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     6301 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     6361 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     6421 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     6481 gtccctcttc tcttatggag atcggccggc cgccggcgtc gacgcgcgcg gccgcggcgc
     6541 gccgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6661 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6721 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6781 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6841 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6901 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6961 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7021 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7081 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7141 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7201 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7261 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7321 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7381 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7441 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7501 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7561 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7621 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7681 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7741 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7801 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7861 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7921 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7981 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8041 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8101 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8161 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8221 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8281 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     8341 ctataaaaat aggcgtatca cgaggccctt tcgtctacgc gcgcgccggc ggcgcgccgt
     8401 cgacggccgg ccgcggccgc
//
