LOCUS       sequence-249388-        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-531    C-Tag Venus210[1] L20 Itgb1 EBPF2
FEATURES             Location/Qualifiers
     source          1..8963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    join(4121..4695,4696..4698,4699..4707)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4288..4305)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4515..4534
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4708..4710,4711..4713,4714..5427)
                     /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(4753..4774)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          5473..5605
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5614..6202
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5667..5687)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6085..6096)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6229..6246)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6235..6346
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6442..6462)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6442..6460)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6481..6497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6481..6497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6494..6516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6599..6618)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6664..6719
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6718..6737)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(7098..7686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7178..7197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7857..8648,8649..8717))
                     /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     8480..8499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8718..8822)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8890..8908
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctgcctcca ggcctcaata ttggccatta gccatattat tcattggtta tatagcataa
       61 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      121 attggctcat gtccaatatg accgccatgt tggcattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggcc
     1021 accatgaatt tgcaactggt ttcctggatt ggattgatca gtttgatttg ttctgtattt
     1081 ggccaaacag ataaaaatag atgtttaaaa gcaaatgcca aatcttgcgg agaatgtata
     1141 caagcagggc caaattgtgg gtggtgtaca aatacgacat ttttgcaaga aggaatgcct
     1201 acttctgcac gatgtgatga tttagaagct ttgaaaaaga agggttgcca gccaagtgac
     1261 atagagaatc ccagaggctc tcaaactata aagaaaaata aaaatgtcac caatcgcagc
     1321 aaagggatgg cagagaagct ccggccagaa gacattactc agatccaacc acaacagctg
     1381 cttctaaaat tgagatcagg agaaccacag aagtttacat taaaattcaa gagggctgaa
     1441 gattacccta ttgatctcta ctaccttatg gatctctcct actctatgaa agatgatctg
     1501 gagaatgtga aaagtcttgg aacggatttg atgaatgaaa tgaggaggat tacttcagac
     1561 ttccgcattg gctttggctc atttgtggag aaaactgtga tgccgtatat tagcacaacc
     1621 ccagcaaagc taagaaatcc ttgtacaagt gaacaaaact gcaccagccc atttagctac
     1681 aaaaatgtgc ttagtcttac tgacagagga gagtttttca atgaacttgt tggtcagcaa
     1741 cgcatatctg gaaacttgga ttctccagaa ggtggctttg atgcaatcat gcaggttgcg
     1801 gtttgtggat cgctgattgg ctggaggaat gtaacacgac tgctggtgtt ttccacggat
     1861 gctgggtttc actttgctgg agatgggaaa cttggtggta ttgttttacc caatgatgga
     1921 caatgtcacc tggaaaataa tgtatataca atgagccatt actatgatta tccttcaatt
     1981 gctcaccttg ttcagaaact aagtgaaaat aatattcaga cgatttttgc agttactgaa
     2041 gagttccaac ctgtttacaa ggaattgaag aatttgattc ctaagtcagc agtgggcaca
     2101 ctgtctggaa actctagtaa tgtgatccag ctaatcatcg atgcctacaa ctctctttct
     2161 tcagaagtca ttctggaaaa tagcaaattg ccagacggag taacaataaa ttacaaatcc
     2221 tattgcaaga atggggtgaa tgggacagga gaaaatggac gaaagtgttc caacatttct
     2281 attggagatg aggttcaatt tgaaattagc ataactgcta ataaatgtcc aaataaggag
     2341 tctgaaacca ttaaaattaa acctctgggc ttcactgaag aagtagaggt cgttcttcag
     2401 ttcatctgta agtgcaattg tcaaagccat ggcatcccag ccagtcccaa gtgccatgag
     2461 ggaaatggga catttgagtg tggagcctgc aggtgcaatg aggggcgtgt tgggaggcac
     2521 tgtgaatgta gcacagatga agtgaacagt gaagacatgg acgcttactg caggaaagag
     2581 aacagttcgg aaatctgcag taataatgga gaatgtgtct gtggacagtg tgtgtgtagg
     2641 aagagagata atacaaatga aatttactct ggaaaattct gcgagtgtga taacttcaac
     2701 tgtgataggt ctaatggctt aatttgtgga ggcaatggcg tgtgcaggtg tcgtgtttgt
     2761 gaatgctatc ccaattacac tggcagtgca tgtgactgtt ctttggacac tggtccatgt
     2821 ctagcgtcaa atggtcagat ctgcaatggc cggggtattt gtgaatgtgg tgcttgtaag
     2881 tgcacagatc ccaagtttca agggccaact tgtgagacat gtcagacctg ccttggcgtc
     2941 tgtgcagagc ataaagaatg tgttcagtgc agagccttca ataaaggaga aaagaaagac
     3001 acgtgtgcac aggagtgctc ccacttcaat ctcaccaaag tagaaagcag ggagaagttg
     3061 ccccagccgg tgcaggtcga tcctgtgacc cattgcaagg agaaggacat tgatgactgc
     3121 tggttctatt tcacctattc agtgaatggc aacaatgaag ctatcgtgca tgttgtggag
     3181 actccagact gtcctactgg tcccgacatc atcccaattg tagcaggcgt ggttgctgga
     3241 attgttctta ttggccttgc cttgctgctg atttggaaac ttttaatgat aattcatgac
     3301 agaagggaat ttgctaaatt tgaaaaggag aaaatgaatg ccaagtggga cacgggtgaa
     3361 aatcctattt acaagagtgc cgtgacaact gtggtcaatc cgaagtatga gggaaaaggg
     3421 agtcctggcg gtggatccgg aggctctggg agctctgcca gcggtggcag caccagtgtg
     3481 tcaaagggag aagagttgtt taccggtgtt gttcccatac tggtggaact ggatggtgac
     3541 gtcaatggac acaagtttag cgtttctggc gaaggagaag gcgatgctac ctatggcaaa
     3601 ctgaccctca aactgatctg tacgactgga aagttgcctg taccttggcc aacactggtc
     3661 acaaccttgg gctatgggct gcaatgcttt gcccgttatc ctgatcacat gaagcaacat
     3721 gacttcttta agtctgctat gccagaggga tatgtgcagg aacggactat cttcttcaag
     3781 gacgacggca actacaagac gagagctgag gtgaaatttg agggtgacac actggtgaat
     3841 cgcatagagc tgaaaggcat cgacttcaaa gaagatggga atattctcgg gcataaactc
     3901 gagtacaact acaattccca caacgtgtac atcactgccg ataagcagaa gaatgggatt
     3961 aaggcgaact tcaagatccg acacaacatc gaggatggtg gggtccagct tgccgaccat
     4021 tatcagcaga acacacccat tggggatgga ccagtcctgt tgccggataa ccactacctc
     4081 agttaccaga gcaaactgtc caaataagaa ttcagatccg cccctctccc tccccccccc
     4141 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     4201 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     4261 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     4321 tcgtgaagga agcagttcct ctggtagctt cttgaagaca aacaacgtct gtagcgaccc
     4381 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     4441 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     4501 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     4561 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
     4621 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     4681 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagct gttcaccggg
     4741 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgagg
     4801 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc
     4861 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgagccacgg cgtgcagtgc
     4921 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4981 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcacctacaa gacccgcgcc
     5041 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg cgtcgacttc
     5101 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaacatc
     5161 tatatcatgg ccgtcaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     5221 gtggaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     5281 ggccccgtgc tgctgcccga cagccactac ctgagcaccc agtccgtgct gagcaaagac
     5341 cccaacgaga agcgcgatca catggtcctg ctggagttcc gcaccgccgc cgggatcact
     5401 ctcggcatgg acgagctgta caagtaaacc ggtgctcgag gtaccgatga atagctaagg
     5461 tcgaggccgc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
     5521 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
     5581 atccactttg cctttctctc cacaggtgtc gacaatcaac ctctggatta caaaatttgt
     5641 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5701 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5761 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5821 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5881 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5941 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6001 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     6061 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6121 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6181 tccctttggg ccgcctcccc gcctggaatt cgagctcggt acgatcagcc tcgactgtgc
     6241 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     6301 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     6361 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     6421 acaatagccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     6481 gtcatagctg tttcctgtgt gaaattgtta tccgctaatt cactcctcag gtgcaggctg
     6541 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     6601 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     6661 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     6721 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     6781 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     6841 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     6901 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     6961 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     7021 ctgtccctct tctcttatgg agatcggccg gccgccggcg tcgacgcgcg cggccgcggc
     7081 gcgccgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8821 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     8881 acctataaaa ataggcgtat cacgaggccc tttcgtctac gcgcgcgccg gcggcgcgcc
     8941 gtcgacggcc ggccgcggcc gcg
//
