LOCUS       sequence-249415-        8435 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 8435)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8435)
  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-510    N-Tag GFP[2] L32 Itgb1 EBPF2
FEATURES             Location/Qualifiers
     source          1..8435
                     /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(3594..4168,4169..4171,4172..4180)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3761..3778)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3988..4007
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4181..4183,4184..4186,4187..4900)
                     /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(4226..4247)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          4946..5078
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5087..5675
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5140..5160)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5558..5569)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5702..5719)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5708..5819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5915..5935)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5915..5933)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5954..5970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5954..5970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5967..5989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6072..6091)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6137..6192
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6191..6210)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(6571..7159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6651..6670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7330..8121,8122..8190))
                     /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     7953..7972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8191..8295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8363..8381
                     /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 gacgtgggtg gcggagggag cgaaggcggc ggatccggag ggcctggctc
     1141 tggcggagag gggagcgctg gaggcgggtc agccggtggt gggagtaatt tgcaactggt
     1201 ttcctggatt ggattgatca gtttgatttg ttctgtattt ggccaaacag ataaaaatag
     1261 atgtttaaaa gcaaatgcca aatcttgcgg agaatgtata caagcagggc caaattgtgg
     1321 gtggtgtaca aatacgacat ttttgcaaga aggaatgcct acttctgcac gatgtgatga
     1381 tttagaagct ttgaaaaaga agggttgcca gccaagtgac atagagaatc ccagaggctc
     1441 tcaaactata aagaaaaata aaaatgtcac caatcgcagc aaagggatgg cagagaagct
     1501 ccggccagaa gacattactc agatccaacc acaacagctg cttctaaaat tgagatcagg
     1561 agaaccacag aagtttacat taaaattcaa gagggctgaa gattacccta ttgatctcta
     1621 ctaccttatg gatctctcct actctatgaa agatgatctg gagaatgtga aaagtcttgg
     1681 aacggatttg atgaatgaaa tgaggaggat tacttcagac ttccgcattg gctttggctc
     1741 atttgtggag aaaactgtga tgccgtatat tagcacaacc ccagcaaagc taagaaatcc
     1801 ttgtacaagt gaacaaaact gcaccagccc atttagctac aaaaatgtgc ttagtcttac
     1861 tgacagagga gagtttttca atgaacttgt tggtcagcaa cgcatatctg gaaacttgga
     1921 ttctccagaa ggtggctttg atgcaatcat gcaggttgcg gtttgtggat cgctgattgg
     1981 ctggaggaat gtaacacgac tgctggtgtt ttccacggat gctgggtttc actttgctgg
     2041 agatgggaaa cttggtggta ttgttttacc caatgatgga caatgtcacc tggaaaataa
     2101 tgtatataca atgagccatt actatgatta tccttcaatt gctcaccttg ttcagaaact
     2161 aagtgaaaat aatattcaga cgatttttgc agttactgaa gagttccaac ctgtttacaa
     2221 ggaattgaag aatttgattc ctaagtcagc agtgggcaca ctgtctggaa actctagtaa
     2281 tgtgatccag ctaatcatcg atgcctacaa ctctctttct tcagaagtca ttctggaaaa
     2341 tagcaaattg ccagacggag taacaataaa ttacaaatcc tattgcaaga atggggtgaa
     2401 tgggacagga gaaaatggac gaaagtgttc caacatttct attggagatg aggttcaatt
     2461 tgaaattagc ataactgcta ataaatgtcc aaataaggag tctgaaacca ttaaaattaa
     2521 acctctgggc ttcactgaag aagtagaggt cgttcttcag ttcatctgta agtgcaattg
     2581 tcaaagccat ggcatcccag ccagtcccaa gtgccatgag ggaaatggga catttgagtg
     2641 tggagcctgc aggtgcaatg aggggcgtgt tgggaggcac tgtgaatgta gcacagatga
     2701 agtgaacagt gaagacatgg acgcttactg caggaaagag aacagttcgg aaatctgcag
     2761 taataatgga gaatgtgtct gtggacagtg tgtgtgtagg aagagagata atacaaatga
     2821 aatttactct ggaaaattct gcgagtgtga taacttcaac tgtgataggt ctaatggctt
     2881 aatttgtgga ggcaatggcg tgtgcaggtg tcgtgtttgt gaatgctatc ccaattacac
     2941 tggcagtgca tgtgactgtt ctttggacac tggtccatgt ctagcgtcaa atggtcagat
     3001 ctgcaatggc cggggtattt gtgaatgtgg tgcttgtaag tgcacagatc ccaagtttca
     3061 agggccaact tgtgagacat gtcagacctg ccttggcgtc tgtgcagagc ataaagaatg
     3121 tgttcagtgc agagccttca ataaaggaga aaagaaagac acgtgtgcac aggagtgctc
     3181 ccacttcaat ctcaccaaag tagaaagcag ggagaagttg ccccagccgg tgcaggtcga
     3241 tcctgtgacc cattgcaagg agaaggacat tgatgactgc tggttctatt tcacctattc
     3301 agtgaatggc aacaatgaag ctatcgtgca tgttgtggag actccagact gtcctactgg
     3361 tcccgacatc atcccaattg tagcaggcgt ggttgctgga attgttctta ttggccttgc
     3421 cttgctgctg atttggaaac ttttaatgat aattcatgac agaagggaat ttgctaaatt
     3481 tgaaaaggag aaaatgaatg ccaagtggga cacgggtgaa aatcctattt acaagagtgc
     3541 cgtgacaact gtggtcaatc cgaagtatga gggaaaataa gaattcagat ccgcccctct
     3601 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     3661 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     3721 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     3781 ggtctgttga atgtcgtgaa ggaagcagtt cctctggtag cttcttgaag acaaacaacg
     3841 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3901 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     3961 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     4021 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc
     4081 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     4141 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     4201 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     4261 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     4321 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
     4381 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4441 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
     4501 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4561 gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     4621 cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa
     4681 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4741 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
     4801 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc
     4861 cgccgggatc actctcggca tggacgagct gtacaagtaa accggtgctc gaggtaccga
     4921 tgaatagcta aggtcgaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga
     4981 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     5041 tggtcttact gacatccact ttgcctttct ctccacaggt gtcgacaatc aacctctgga
     5101 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5161 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5221 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5281 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5341 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5401 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5461 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     5521 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5581 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5641 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc ggtacgatca
     5701 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     5761 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     5821 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     5881 gaggattggg aagacaatag cccagctttt gttcccttta gtgagggtta attgcgcgct
     5941 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgcta attcactcct
     6001 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     6061 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     6121 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     6181 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     6241 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     6301 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     6361 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     6421 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     6481 ctcccagtca tagctgtccc tcttctctta tggagatcgg ccggccgccg gcgtcgacgc
     6541 gcgcggccgc ggcgcgccgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6601 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6661 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6721 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6781 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6841 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6901 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6961 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7021 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7081 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7321 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7381 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7441 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7501 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7561 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7621 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7681 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7741 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7801 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7861 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7921 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7981 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8041 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8101 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8161 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8221 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8281 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8341 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tacgcgcgcg
     8401 ccggcggcgc gccgtcgacg gccggccgcg gccgc
//
