LOCUS       sequence-249411-        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-506    N-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"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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 caccatggaa aagagggacc acatggtgct gctcgagtac gtcacagcag ctggcatcac
     1081 cgatgccagc gggagctctg ccagcggtgg cagcacctcc aatttgcaac tggtttcctg
     1141 gattggattg atcagtttga tttgttctgt atttggccaa acagataaaa atagatgttt
     1201 aaaagcaaat gccaaatctt gcggagaatg tatacaagca gggccaaatt gtgggtggtg
     1261 tacaaatacg acatttttgc aagaaggaat gcctacttct gcacgatgtg atgatttaga
     1321 agctttgaaa aagaagggtt gccagccaag tgacatagag aatcccagag gctctcaaac
     1381 tataaagaaa aataaaaatg tcaccaatcg cagcaaaggg atggcagaga agctccggcc
     1441 agaagacatt actcagatcc aaccacaaca gctgcttcta aaattgagat caggagaacc
     1501 acagaagttt acattaaaat tcaagagggc tgaagattac cctattgatc tctactacct
     1561 tatggatctc tcctactcta tgaaagatga tctggagaat gtgaaaagtc ttggaacgga
     1621 tttgatgaat gaaatgagga ggattacttc agacttccgc attggctttg gctcatttgt
     1681 ggagaaaact gtgatgccgt atattagcac aaccccagca aagctaagaa atccttgtac
     1741 aagtgaacaa aactgcacca gcccatttag ctacaaaaat gtgcttagtc ttactgacag
     1801 aggagagttt ttcaatgaac ttgttggtca gcaacgcata tctggaaact tggattctcc
     1861 agaaggtggc tttgatgcaa tcatgcaggt tgcggtttgt ggatcgctga ttggctggag
     1921 gaatgtaaca cgactgctgg tgttttccac ggatgctggg tttcactttg ctggagatgg
     1981 gaaacttggt ggtattgttt tacccaatga tggacaatgt cacctggaaa ataatgtata
     2041 tacaatgagc cattactatg attatccttc aattgctcac cttgttcaga aactaagtga
     2101 aaataatatt cagacgattt ttgcagttac tgaagagttc caacctgttt acaaggaatt
     2161 gaagaatttg attcctaagt cagcagtggg cacactgtct ggaaactcta gtaatgtgat
     2221 ccagctaatc atcgatgcct acaactctct ttcttcagaa gtcattctgg aaaatagcaa
     2281 attgccagac ggagtaacaa taaattacaa atcctattgc aagaatgggg tgaatgggac
     2341 aggagaaaat ggacgaaagt gttccaacat ttctattgga gatgaggttc aatttgaaat
     2401 tagcataact gctaataaat gtccaaataa ggagtctgaa accattaaaa ttaaacctct
     2461 gggcttcact gaagaagtag aggtcgttct tcagttcatc tgtaagtgca attgtcaaag
     2521 ccatggcatc ccagccagtc ccaagtgcca tgagggaaat gggacatttg agtgtggagc
     2581 ctgcaggtgc aatgaggggc gtgttgggag gcactgtgaa tgtagcacag atgaagtgaa
     2641 cagtgaagac atggacgctt actgcaggaa agagaacagt tcggaaatct gcagtaataa
     2701 tggagaatgt gtctgtggac agtgtgtgtg taggaagaga gataatacaa atgaaattta
     2761 ctctggaaaa ttctgcgagt gtgataactt caactgtgat aggtctaatg gcttaatttg
     2821 tggaggcaat ggcgtgtgca ggtgtcgtgt ttgtgaatgc tatcccaatt acactggcag
     2881 tgcatgtgac tgttctttgg acactggtcc atgtctagcg tcaaatggtc agatctgcaa
     2941 tggccggggt atttgtgaat gtggtgcttg taagtgcaca gatcccaagt ttcaagggcc
     3001 aacttgtgag acatgtcaga cctgccttgg cgtctgtgca gagcataaag aatgtgttca
     3061 gtgcagagcc ttcaataaag gagaaaagaa agacacgtgt gcacaggagt gctcccactt
     3121 caatctcacc aaagtagaaa gcagggagaa gttgccccag ccggtgcagg tcgatcctgt
     3181 gacccattgc aaggagaagg acattgatga ctgctggttc tatttcacct attcagtgaa
     3241 tggcaacaat gaagctatcg tgcatgttgt ggagactcca gactgtccta ctggtcccga
     3301 catcatccca attgtagcag gcgtggttgc tggaattgtt cttattggcc ttgccttgct
     3361 gctgatttgg aaacttttaa tgataattca tgacagaagg gaatttgcta aatttgaaaa
     3421 ggagaaaatg aatgccaagt gggacacggg tgaaaatcct atttacaaga gtgccgtgac
     3481 aactgtggtc aatccgaagt atgagggaaa ataagaattc 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 gctaaggtag 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
//
