LOCUS       sequence-249387-        8933 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 8933)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8933)
  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-530    N-Tag Venus210[1] L10 Itgb1 EBPF2
FEATURES             Location/Qualifiers
     source          1..8933
                     /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(4092..4666,4667..4669,4670..4678)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4259..4276)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4486..4505
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4679..4681,4682..4684,4685..5398)
                     /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(4724..4745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          5444..5576
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5585..6173
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5638..5658)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6056..6067)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6200..6217)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6206..6317
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6413..6433)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6413..6431)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6452..6468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6452..6468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6465..6487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6570..6589)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6635..6690
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6689..6708)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(7069..7657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7149..7168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7828..8619,8620..8688))
                     /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     8451..8470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8689..8793)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8861..8879
                     /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 caccatggtg tcaaagggag aagagttgtt taccggtgtt gttcccatac tggtggaact
     1081 ggatggtgac gtcaatggac acaagtttag cgtttctggc gaaggagaag gcgatgctac
     1141 ctatggcaaa ctgaccctca aactgatctg tacgactgga aagttgcctg taccttggcc
     1201 aacactggtc acaaccttgg gctatgggct gcaatgcttt gcccgttatc ctgatcacat
     1261 gaagcaacat gacttcttta agtctgctat gccagaggga tatgtgcagg aacggactat
     1321 cttcttcaag gacgacggca actacaagac gagagctgag gtgaaatttg agggtgacac
     1381 actggtgaat cgcatagagc tgaaaggcat cgacttcaaa gaagatggga atattctcgg
     1441 gcataaactc gagtacaact acaattccca caacgtgtac atcactgccg ataagcagaa
     1501 gaatgggatt aaggcgaact tcaagatccg acacaacatc gaggatggtg gggtccagct
     1561 tgccgaccat tatcagcaga acacacccat tggggatgga ccagtcctgt tgccggataa
     1621 ccactacctc agttaccaga gcaaactgtc caaagggagc tctgccagcg gtggcagcac
     1681 ctccaatttg caactggttt cctggattgg attgatcagt ttgatttgtt ctgtatttgg
     1741 ccaaacagat aaaaatagat gtttaaaagc aaatgccaaa tcttgcggag aatgtataca
     1801 agcagggcca aattgtgggt ggtgtacaaa tacgacattt ttgcaagaag gaatgcctac
     1861 ttctgcacga tgtgatgatt tagaagcttt gaaaaagaag ggttgccagc caagtgacat
     1921 agagaatccc agaggctctc aaactataaa gaaaaataaa aatgtcacca atcgcagcaa
     1981 agggatggca gagaagctcc ggccagaaga cattactcag atccaaccac aacagctgct
     2041 tctaaaattg agatcaggag aaccacagaa gtttacatta aaattcaaga gggctgaaga
     2101 ttaccctatt gatctctact accttatgga tctctcctac tctatgaaag atgatctgga
     2161 gaatgtgaaa agtcttggaa cggatttgat gaatgaaatg aggaggatta cttcagactt
     2221 ccgcattggc tttggctcat ttgtggagaa aactgtgatg ccgtatatta gcacaacccc
     2281 agcaaagcta agaaatcctt gtacaagtga acaaaactgc accagcccat ttagctacaa
     2341 aaatgtgctt agtcttactg acagaggaga gtttttcaat gaacttgttg gtcagcaacg
     2401 catatctgga aacttggatt ctccagaagg tggctttgat gcaatcatgc aggttgcggt
     2461 ttgtggatcg ctgattggct ggaggaatgt aacacgactg ctggtgtttt ccacggatgc
     2521 tgggtttcac tttgctggag atgggaaact tggtggtatt gttttaccca atgatggaca
     2581 atgtcacctg gaaaataatg tatatacaat gagccattac tatgattatc cttcaattgc
     2641 tcaccttgtt cagaaactaa gtgaaaataa tattcagacg atttttgcag ttactgaaga
     2701 gttccaacct gtttacaagg aattgaagaa tttgattcct aagtcagcag tgggcacact
     2761 gtctggaaac tctagtaatg tgatccagct aatcatcgat gcctacaact ctctttcttc
     2821 agaagtcatt ctggaaaata gcaaattgcc agacggagta acaataaatt acaaatccta
     2881 ttgcaagaat ggggtgaatg ggacaggaga aaatggacga aagtgttcca acatttctat
     2941 tggagatgag gttcaatttg aaattagcat aactgctaat aaatgtccaa ataaggagtc
     3001 tgaaaccatt aaaattaaac ctctgggctt cactgaagaa gtagaggtcg ttcttcagtt
     3061 catctgtaag tgcaattgtc aaagccatgg catcccagcc agtcccaagt gccatgaggg
     3121 aaatgggaca tttgagtgtg gagcctgcag gtgcaatgag gggcgtgttg ggaggcactg
     3181 tgaatgtagc acagatgaag tgaacagtga agacatggac gcttactgca ggaaagagaa
     3241 cagttcggaa atctgcagta ataatggaga atgtgtctgt ggacagtgtg tgtgtaggaa
     3301 gagagataat acaaatgaaa tttactctgg aaaattctgc gagtgtgata acttcaactg
     3361 tgataggtct aatggcttaa tttgtggagg caatggcgtg tgcaggtgtc gtgtttgtga
     3421 atgctatccc aattacactg gcagtgcatg tgactgttct ttggacactg gtccatgtct
     3481 agcgtcaaat ggtcagatct gcaatggccg gggtatttgt gaatgtggtg cttgtaagtg
     3541 cacagatccc aagtttcaag ggccaacttg tgagacatgt cagacctgcc ttggcgtctg
     3601 tgcagagcat aaagaatgtg ttcagtgcag agccttcaat aaaggagaaa agaaagacac
     3661 gtgtgcacag gagtgctccc acttcaatct caccaaagta gaaagcaggg agaagttgcc
     3721 ccagccggtg caggtcgatc ctgtgaccca ttgcaaggag aaggacattg atgactgctg
     3781 gttctatttc acctattcag tgaatggcaa caatgaagct atcgtgcatg ttgtggagac
     3841 tccagactgt cctactggtc ccgacatcat cccaattgta gcaggcgtgg ttgctggaat
     3901 tgttcttatt ggccttgcct tgctgctgat ttggaaactt ttaatgataa ttcatgacag
     3961 aagggaattt gctaaatttg aaaaggagaa aatgaatgcc aagtgggaca cgggtgaaaa
     4021 tcctatttac aagagtgccg tgacaactgt ggtcaatccg aagtatgagg gaaaataaga
     4081 attcagatcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     4141 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     4201 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     4261 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggtagct
     4321 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     4381 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     4441 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     4501 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     4561 gggcctcggt acacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     4621 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     4681 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4741 cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc gagggcgatg ccaccaacgg
     4801 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4861 cgtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     4921 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4981 caaggacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     5041 gaaccgcatc gagctgaagg gcgtcgactt caaggaggac ggcaacatcc tggggcacaa
     5101 gctggagtac aacttcaaca gccacaacat ctatatcatg gccgtcaagc agaagaacgg
     5161 catcaaggtg aacttcaaga tccgccacaa cgtggaggac ggcagcgtgc agctcgccga
     5221 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acagccacta
     5281 cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5341 gctggagttc cgcaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaac
     5401 cggtgctcga ggtaccgatg aatagctaag gtagaggccg caggtaagta tcaaggttac
     5461 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
     5521 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     5581 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5641 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5701 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5761 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5821 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5881 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5941 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     6001 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6061 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     6121 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     6181 tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     6241 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     6301 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     6361 gggcaggaca gcaaggggga ggattgggaa gacaatagcc cagcttttgt tccctttagt
     6421 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6481 atccgctaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     6541 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     6601 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     6661 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     6721 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     6781 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     6841 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     6901 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     6961 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatcggcc
     7021 ggccgccggc gtcgacgcgc gcggccgcgg cgcgccgttg ctggcgtttt tccataggct
     7081 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7141 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7201 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7261 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7321 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7381 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7441 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7501 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7561 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7621 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7681 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7741 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7801 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7861 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7921 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7981 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8041 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8101 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8161 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8221 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8281 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8341 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8401 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8461 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8521 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8581 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8641 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8701 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8761 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8821 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8881 ctttcgtcta cgcgcgcgcc ggcggcgcgc cgtcgacggc cggccgcggc cgc
//
