LOCUS       sequence-270546-        8369 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 8369)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8369)
  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-505    C-Tag GFP[2] L10 Itgb1 EBPF2
FEATURES             Location/Qualifiers
     source          1..8369
                     /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"
     misc_feature    join(3528..4102,4103..4105,4106..4114)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3695..3712)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3922..3941
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4115..4117,4118..4120,4121..4834)
                     /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(4160..4181)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          4880..5012
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5021..5609
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5074..5094)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5492..5503)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5636..5653)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5642..5753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5849..5869)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5849..5867)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5888..5904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5888..5904)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5901..5923)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6006..6025)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6071..6126
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6125..6144)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6505..7093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6585..6604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7264..8055,8056..8124))
                     /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     7887..7906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8125..8229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8297..8315
                     /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 caccatgaat ttgcaactgg tttcctggat tggattgatc agtttgattt gttctgtatt
     1081 tggccaaaca gataaaaata gatgtttaaa agcaaatgcc aaatcttgcg gagaatgtat
     1141 acaagcaggg ccaaattgtg ggtggtgtac aaatacgaca tttttgcaag aaggaatgcc
     1201 tacttctgca cgatgtgatg atttagaagc tttgaaaaag aagggttgcc agccaagtga
     1261 catagagaat cccagaggct ctcaaactat aaagaaaaat aaaaatgtca ccaatcgcag
     1321 caaagggatg gcagagaagc tccggccaga agacattact cagatccaac cacaacagct
     1381 gcttctaaaa ttgagatcag gagaaccaca gaagtttaca ttaaaattca agagggctga
     1441 agattaccct attgatctct actaccttat ggatctctcc tactctatga aagatgatct
     1501 ggagaatgtg aaaagtcttg gaacggattt gatgaatgaa atgaggagga ttacttcaga
     1561 cttccgcatt ggctttggct catttgtgga gaaaactgtg atgccgtata ttagcacaac
     1621 cccagcaaag ctaagaaatc cttgtacaag tgaacaaaac tgcaccagcc catttagcta
     1681 caaaaatgtg cttagtctta ctgacagagg agagtttttc aatgaacttg ttggtcagca
     1741 acgcatatct ggaaacttgg attctccaga aggtggcttt gatgcaatca tgcaggttgc
     1801 ggtttgtgga tcgctgattg gctggaggaa tgtaacacga ctgctggtgt tttccacgga
     1861 tgctgggttt cactttgctg gagatgggaa acttggtggt attgttttac ccaatgatgg
     1921 acaatgtcac ctggaaaata atgtatatac aatgagccat tactatgatt atccttcaat
     1981 tgctcacctt gttcagaaac taagtgaaaa taatattcag acgatttttg cagttactga
     2041 agagttccaa cctgtttaca aggaattgaa gaatttgatt cctaagtcag cagtgggcac
     2101 actgtctgga aactctagta atgtgatcca gctaatcatc gatgcctaca actctctttc
     2161 ttcagaagtc attctggaaa atagcaaatt gccagacgga gtaacaataa attacaaatc
     2221 ctattgcaag aatggggtga atgggacagg agaaaatgga cgaaagtgtt ccaacatttc
     2281 tattggagat gaggttcaat ttgaaattag cataactgct aataaatgtc caaataagga
     2341 gtctgaaacc attaaaatta aacctctggg cttcactgaa gaagtagagg tcgttcttca
     2401 gttcatctgt aagtgcaatt gtcaaagcca tggcatccca gccagtccca agtgccatga
     2461 gggaaatggg acatttgagt gtggagcctg caggtgcaat gaggggcgtg ttgggaggca
     2521 ctgtgaatgt agcacagatg aagtgaacag tgaagacatg gacgcttact gcaggaaaga
     2581 gaacagttcg gaaatctgca gtaataatgg agaatgtgtc tgtggacagt gtgtgtgtag
     2641 gaagagagat aatacaaatg aaatttactc tggaaaattc tgcgagtgtg ataacttcaa
     2701 ctgtgatagg tctaatggct taatttgtgg aggcaatggc gtgtgcaggt gtcgtgtttg
     2761 tgaatgctat cccaattaca ctggcagtgc atgtgactgt tctttggaca ctggtccatg
     2821 tctagcgtca aatggtcaga tctgcaatgg ccggggtatt tgtgaatgtg gtgcttgtaa
     2881 gtgcacagat cccaagtttc aagggccaac ttgtgagaca tgtcagacct gccttggcgt
     2941 ctgtgcagag cataaagaat gtgttcagtg cagagccttc aataaaggag aaaagaaaga
     3001 cacgtgtgca caggagtgct cccacttcaa tctcaccaaa gtagaaagca gggagaagtt
     3061 gccccagccg gtgcaggtcg atcctgtgac ccattgcaag gagaaggaca ttgatgactg
     3121 ctggttctat ttcacctatt cagtgaatgg caacaatgaa gctatcgtgc atgttgtgga
     3181 gactccagac tgtcctactg gtcccgacat catcccaatt gtagcaggcg tggttgctgg
     3241 aattgttctt attggccttg ccttgctgct gatttggaaa cttttaatga taattcatga
     3301 cagaagggaa tttgctaaat ttgaaaagga gaaaatgaat gccaagtggg acacgggtga
     3361 aaatcctatt tacaagagtg ccgtgacaac tgtggtcaat ccgaagtatg agggaaaagg
     3421 gagctctgcc agcggtggca gcacctccga aaagagggac cacatggtgc tgctcgagta
     3481 cgtcacagca gctggcatca ccgatgccag ctaagaattc agatccgccc ctctccctcc
     3541 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3601 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3661 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3721 ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt gaagacaaac aacgtctgta
     3781 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3841 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3901 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3961 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     4021 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4081 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt
     4141 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4201 cgtgaggggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
     4261 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt
     4321 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4381 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca cctacaagac
     4441 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcgt
     4501 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca
     4561 caacatctat atcatggccg tcaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4621 ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4681 cggcgacggc cccgtgctgc tgcccgacag ccactacctg agcacccagt ccgtgctgag
     4741 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttccgca ccgccgccgg
     4801 gatcactctc ggcatggacg agctgtacaa gtaaaccggt gctcgaggta ccgatgaata
     4861 gctaaggtcg aggccgcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     4921 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     4981 tactgacatc cactttgcct ttctctccac aggtgtcgac aatcaacctc tggattacaa
     5041 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5101 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5161 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5221 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5281 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5341 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5401 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     5461 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5521 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5581 tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacg atcagcctcg
     5641 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5701 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5761 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5821 tgggaagaca atagcccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     5881 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaattcac tcctcaggtg
     5941 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     6001 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     6061 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     6121 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     6181 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     6241 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     6301 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     6361 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     6421 gtcatagctg tccctcttct cttatggaga tcggccggcc gccggcgtcg acgcgcgcgg
     6481 ccgcggcgcg ccgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     8281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctacgcg cgcgccggcg
     8341 gcgcgccgtc gacggccggc cgcggccgc
//
