LOCUS       sequence-249389-        8963 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 8963)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8963)
  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-532    N-Tag Venus210[1] L20 Itgb1 EBPF2
FEATURES             Location/Qualifiers
     source          1..8963
                     /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(4122..4696,4697..4699,4700..4708)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4289..4306)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4516..4535
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4709..4711,4712..4714,4715..5428)
                     /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(4754..4775)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          5474..5606
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5615..6203
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5668..5688)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6086..6097)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6230..6247)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6236..6347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6443..6463)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6443..6461)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6482..6498)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6482..6498)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6495..6517)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6600..6619)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6665..6720
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6719..6738)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(7099..7687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7179..7198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7858..8649,8650..8718))
                     /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     8481..8500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8719..8823)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8891..8909
                     /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 caaagggagt cctggcggtg gatccggagg
     1681 ctctgggagc tctgccagcg gtggcagcac ctccaatttg caactggttt cctggattgg
     1741 attgatcagt ttgatttgtt ctgtatttgg ccaaacagat aaaaatagat gtttaaaagc
     1801 aaatgccaaa tcttgcggag aatgtataca agcagggcca aattgtgggt ggtgtacaaa
     1861 tacgacattt ttgcaagaag gaatgcctac ttctgcacga tgtgatgatt tagaagcttt
     1921 gaaaaagaag ggttgccagc caagtgacat agagaatccc agaggctctc aaactataaa
     1981 gaaaaataaa aatgtcacca atcgcagcaa agggatggca gagaagctcc ggccagaaga
     2041 cattactcag atccaaccac aacagctgct tctaaaattg agatcaggag aaccacagaa
     2101 gtttacatta aaattcaaga gggctgaaga ttaccctatt gatctctact accttatgga
     2161 tctctcctac tctatgaaag atgatctgga gaatgtgaaa agtcttggaa cggatttgat
     2221 gaatgaaatg aggaggatta cttcagactt ccgcattggc tttggctcat ttgtggagaa
     2281 aactgtgatg ccgtatatta gcacaacccc agcaaagcta agaaatcctt gtacaagtga
     2341 acaaaactgc accagcccat ttagctacaa aaatgtgctt agtcttactg acagaggaga
     2401 gtttttcaat gaacttgttg gtcagcaacg catatctgga aacttggatt ctccagaagg
     2461 tggctttgat gcaatcatgc aggttgcggt ttgtggatcg ctgattggct ggaggaatgt
     2521 aacacgactg ctggtgtttt ccacggatgc tgggtttcac tttgctggag atgggaaact
     2581 tggtggtatt gttttaccca atgatggaca atgtcacctg gaaaataatg tatatacaat
     2641 gagccattac tatgattatc cttcaattgc tcaccttgtt cagaaactaa gtgaaaataa
     2701 tattcagacg atttttgcag ttactgaaga gttccaacct gtttacaagg aattgaagaa
     2761 tttgattcct aagtcagcag tgggcacact gtctggaaac tctagtaatg tgatccagct
     2821 aatcatcgat gcctacaact ctctttcttc agaagtcatt ctggaaaata gcaaattgcc
     2881 agacggagta acaataaatt acaaatccta ttgcaagaat ggggtgaatg ggacaggaga
     2941 aaatggacga aagtgttcca acatttctat tggagatgag gttcaatttg aaattagcat
     3001 aactgctaat aaatgtccaa ataaggagtc tgaaaccatt aaaattaaac ctctgggctt
     3061 cactgaagaa gtagaggtcg ttcttcagtt catctgtaag tgcaattgtc aaagccatgg
     3121 catcccagcc agtcccaagt gccatgaggg aaatgggaca tttgagtgtg gagcctgcag
     3181 gtgcaatgag gggcgtgttg ggaggcactg tgaatgtagc acagatgaag tgaacagtga
     3241 agacatggac gcttactgca ggaaagagaa cagttcggaa atctgcagta ataatggaga
     3301 atgtgtctgt ggacagtgtg tgtgtaggaa gagagataat acaaatgaaa tttactctgg
     3361 aaaattctgc gagtgtgata acttcaactg tgataggtct aatggcttaa tttgtggagg
     3421 caatggcgtg tgcaggtgtc gtgtttgtga atgctatccc aattacactg gcagtgcatg
     3481 tgactgttct ttggacactg gtccatgtct agcgtcaaat ggtcagatct gcaatggccg
     3541 gggtatttgt gaatgtggtg cttgtaagtg cacagatccc aagtttcaag ggccaacttg
     3601 tgagacatgt cagacctgcc ttggcgtctg tgcagagcat aaagaatgtg ttcagtgcag
     3661 agccttcaat aaaggagaaa agaaagacac gtgtgcacag gagtgctccc acttcaatct
     3721 caccaaagta gaaagcaggg agaagttgcc ccagccggtg caggtcgatc ctgtgaccca
     3781 ttgcaaggag aaggacattg atgactgctg gttctatttc acctattcag tgaatggcaa
     3841 caatgaagct atcgtgcatg ttgtggagac tccagactgt cctactggtc ccgacatcat
     3901 cccaattgta gcaggcgtgg ttgctggaat tgttcttatt ggccttgcct tgctgctgat
     3961 ttggaaactt ttaatgataa ttcatgacag aagggaattt gctaaatttg aaaaggagaa
     4021 aatgaatgcc aagtgggaca cgggtgaaaa tcctatttac aagagtgccg tgacaactgt
     4081 ggtcaatccg aagtatgagg gaaaataaga attcagatcc gcccctctcc ctcccccccc
     4141 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     4201 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     4261 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     4321 gtcgtgaagg aagcagttcc tctggtagct tcttgaagac aaacaacgtc tgtagcgacc
     4381 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     4441 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     4501 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     4561 aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt
     4621 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     4681 aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg
     4741 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgag
     4801 gggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac
     4861 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagccacg gcgtgcagtg
     4921 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4981 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcacctaca agacccgcgc
     5041 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcgtcgactt
     5101 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacat
     5161 ctatatcatg gccgtcaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     5221 cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     5281 cggccccgtg ctgctgcccg acagccacta cctgagcacc cagtccgtgc tgagcaaaga
     5341 ccccaacgag aagcgcgatc acatggtcct gctggagttc cgcaccgccg ccgggatcac
     5401 tctcggcatg gacgagctgt acaagtaaac cggtgctcga ggtaccgatg aatagctaag
     5461 gtagaggccg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     5521 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     5581 catccacttt gcctttctct ccacaggtgt cgacaatcaa cctctggatt acaaaatttg
     5641 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5701 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5761 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5821 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5881 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5941 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6001 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     6061 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6121 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6181 ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacgatcagc ctcgactgtg
     6241 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6301 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6361 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6421 gacaatagcc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     6481 ggtcatagct gtttcctgtg tgaaattgtt atccgctaat tcactcctca ggtgcaggct
     6541 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     6601 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     6661 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     6721 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     6781 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     6841 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     6901 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     6961 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     7021 gctgtccctc ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc gcggccgcgg
     7081 cgcgccgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7501 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8821 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8881 aacctataaa aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc ggcggcgcgc
     8941 cgtcgacggc cggccgcggc cgc
//
