LOCUS       sequence-249413-        8399 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 8399)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8399)
  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-508    N-Tag GFP[2] L20 Itgb1 EBPF2
FEATURES             Location/Qualifiers
     source          1..8399
                     /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(3558..4132,4133..4135,4136..4144)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3725..3742)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3952..3971
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4145..4147,4148..4150,4151..4864)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(4190..4211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          4910..5042
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5051..5639
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5104..5124)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5522..5533)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5666..5683)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5672..5783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5879..5899)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5879..5897)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5918..5934)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5918..5934)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5931..5953)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6036..6055)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6101..6156
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6155..6174)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6535..7123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6615..6634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7294..8085,8086..8154))
                     /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     7917..7936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8155..8259)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8327..8345
                     /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 caccatggaa aagagggacc acatggtgct gctcgagtac gtcacagcag ctggcatcac
     1081 cgatgccagc gggagtcctg gcggtggatc cggaggctct gggagctctg ccagcggtgg
     1141 cagcacctcc aatttgcaac tggtttcctg gattggattg atcagtttga tttgttctgt
     1201 atttggccaa acagataaaa atagatgttt aaaagcaaat gccaaatctt gcggagaatg
     1261 tatacaagca gggccaaatt gtgggtggtg tacaaatacg acatttttgc aagaaggaat
     1321 gcctacttct gcacgatgtg atgatttaga agctttgaaa aagaagggtt gccagccaag
     1381 tgacatagag aatcccagag gctctcaaac tataaagaaa aataaaaatg tcaccaatcg
     1441 cagcaaaggg atggcagaga agctccggcc agaagacatt actcagatcc aaccacaaca
     1501 gctgcttcta aaattgagat caggagaacc acagaagttt acattaaaat tcaagagggc
     1561 tgaagattac cctattgatc tctactacct tatggatctc tcctactcta tgaaagatga
     1621 tctggagaat gtgaaaagtc ttggaacgga tttgatgaat gaaatgagga ggattacttc
     1681 agacttccgc attggctttg gctcatttgt ggagaaaact gtgatgccgt atattagcac
     1741 aaccccagca aagctaagaa atccttgtac aagtgaacaa aactgcacca gcccatttag
     1801 ctacaaaaat gtgcttagtc ttactgacag aggagagttt ttcaatgaac ttgttggtca
     1861 gcaacgcata tctggaaact tggattctcc agaaggtggc tttgatgcaa tcatgcaggt
     1921 tgcggtttgt ggatcgctga ttggctggag gaatgtaaca cgactgctgg tgttttccac
     1981 ggatgctggg tttcactttg ctggagatgg gaaacttggt ggtattgttt tacccaatga
     2041 tggacaatgt cacctggaaa ataatgtata tacaatgagc cattactatg attatccttc
     2101 aattgctcac cttgttcaga aactaagtga aaataatatt cagacgattt ttgcagttac
     2161 tgaagagttc caacctgttt acaaggaatt gaagaatttg attcctaagt cagcagtggg
     2221 cacactgtct ggaaactcta gtaatgtgat ccagctaatc atcgatgcct acaactctct
     2281 ttcttcagaa gtcattctgg aaaatagcaa attgccagac ggagtaacaa taaattacaa
     2341 atcctattgc aagaatgggg tgaatgggac aggagaaaat ggacgaaagt gttccaacat
     2401 ttctattgga gatgaggttc aatttgaaat tagcataact gctaataaat gtccaaataa
     2461 ggagtctgaa accattaaaa ttaaacctct gggcttcact gaagaagtag aggtcgttct
     2521 tcagttcatc tgtaagtgca attgtcaaag ccatggcatc ccagccagtc ccaagtgcca
     2581 tgagggaaat gggacatttg agtgtggagc ctgcaggtgc aatgaggggc gtgttgggag
     2641 gcactgtgaa tgtagcacag atgaagtgaa cagtgaagac atggacgctt actgcaggaa
     2701 agagaacagt tcggaaatct gcagtaataa tggagaatgt gtctgtggac agtgtgtgtg
     2761 taggaagaga gataatacaa atgaaattta ctctggaaaa ttctgcgagt gtgataactt
     2821 caactgtgat aggtctaatg gcttaatttg tggaggcaat ggcgtgtgca ggtgtcgtgt
     2881 ttgtgaatgc tatcccaatt acactggcag tgcatgtgac tgttctttgg acactggtcc
     2941 atgtctagcg tcaaatggtc agatctgcaa tggccggggt atttgtgaat gtggtgcttg
     3001 taagtgcaca gatcccaagt ttcaagggcc aacttgtgag acatgtcaga cctgccttgg
     3061 cgtctgtgca gagcataaag aatgtgttca gtgcagagcc ttcaataaag gagaaaagaa
     3121 agacacgtgt gcacaggagt gctcccactt caatctcacc aaagtagaaa gcagggagaa
     3181 gttgccccag ccggtgcagg tcgatcctgt gacccattgc aaggagaagg acattgatga
     3241 ctgctggttc tatttcacct attcagtgaa tggcaacaat gaagctatcg tgcatgttgt
     3301 ggagactcca gactgtccta ctggtcccga catcatccca attgtagcag gcgtggttgc
     3361 tggaattgtt cttattggcc ttgccttgct gctgatttgg aaacttttaa tgataattca
     3421 tgacagaagg gaatttgcta aatttgaaaa ggagaaaatg aatgccaagt gggacacggg
     3481 tgaaaatcct atttacaaga gtgccgtgac aactgtggtc aatccgaagt atgagggaaa
     3541 ataagaattc agatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     3601 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     3661 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     3721 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     3781 gtagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     3841 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     3901 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     3961 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     4021 gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     4081 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     4141 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4201 ggacggcgac gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac
     4261 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4321 caccctcgtg accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat
     4381 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4441 cttcttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac
     4501 cctggtgaac cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg
     4561 gcacaagctg gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa
     4621 gaacggcatc aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct
     4681 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag
     4741 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
     4801 ggtcctgctg gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4861 gtaaaccggt gctcgaggta ccgatgaata gctaaggtag aggccgcagg taagtatcaa
     4921 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     4981 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     5041 aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5101 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5161 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5221 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5281 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5341 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5401 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5461 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5521 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5581 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5641 tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     5701 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     5761 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     5821 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc
     5881 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5941 attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     6001 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     6061 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     6121 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     6181 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     6241 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     6301 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     6361 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     6421 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     6481 tcggccggcc gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca
     6541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8281 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     8341 gaggcccttt cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc cgcggccgc
//
