LOCUS       sequence-249401-        7655 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 7655)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7655)
  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-544    N-Tag Venus210[1] L20 Tln1(rod) E...
FEATURES             Location/Qualifiers
     source          1..7655
                     /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(2814..3388,3389..3391,3392..3400)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2981..2998)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3208..3227
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3401..3403,3404..3406,3407..4120)
                     /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(3446..3467)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          4166..4298
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    4307..4895
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4360..4380)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4778..4789)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4922..4939)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4928..5039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5135..5155)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5135..5153)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5174..5190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5174..5190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5187..5209)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5292..5311)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5357..5412
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5411..5430)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5791..6379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5871..5890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6550..7341,7342..7410))
                     /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     7173..7192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7411..7515)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7583..7601
                     /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 ctccgctgtg tctggtatca ttgctgacct
     1741 cgacaccacc atcatgttcg ctactgctgg cacacttaac cgtgagggtg ctgaaacttt
     1801 tgctgaccac cgggagggta tcttaaagac agcaaaggtt cttgtggagg acaccaaggt
     1861 cctagtgcag aatgcagctg ggagccagga gaagttggca caagccgccc agtcctccgt
     1921 ggccaccatt acccgcctcg ctgatgtggt caagctcggt gcagccagcc taggagccga
     1981 agaccctgaa actcaggtgg tgctgatcaa tgcagtaaag gacgtagcca aggccctggg
     2041 tgacctcatc agcgctacga aggctgcagc gggcaaagtt ggggatgacc ctgcagtgtg
     2101 gcagctcaag aactctgcca aggtgatggt gaccaatgtg acatcattgc tcaagacagt
     2161 gaaggctgtg gaagatgagg ccaccaaagg cacacgggcc ctagaggcaa ccacagagca
     2221 catacgtcag gaactggcgg tcttctgttc cccagagcca cctgccaaga cctctacccc
     2281 tgaagacttc atccgaatga ccaagggtat tactatggca acagccaaag ccgttgctgc
     2341 tggcaattcc tgtcgacagg aagatgtcat tgccacagcc aatctgagcc gacgggctat
     2401 tgcggacatg cttcgggctt gcaaggaagc agctttccac ccagaagtgg cgcctgatgt
     2461 acggctccga gccctgcact atggccggga gtgtgccaat ggttacctgg aactgctgga
     2521 ccacgtgctg ctgacccttc agaagccaaa cccagacctg aagcagcagc taactggaca
     2581 ctcaaagcgg gttgctggct ctgtgactga gctcatccag gctgctgaag ccatgaaagg
     2641 aacagagtgg gtggacccag aggaccctac tgtcattgct gagaatgagc tcttgggagc
     2701 cgcagccgcc atcgaggctg cagccaagaa gctggagcag ctgaagccca gggccaaacc
     2761 caaggaggcg gatgagtcct tgaactttga ggaacaataa gaattcagat ccgcccctct
     2821 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     2881 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     2941 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     3001 ggtctgttga atgtcgtgaa ggaagcagtt cctctggtag cttcttgaag acaaacaacg
     3061 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3121 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     3181 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     3241 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc
     3301 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     3361 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     3421 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3481 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     3541 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
     3601 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3661 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
     3721 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3781 gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     3841 cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa
     3901 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3961 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
     4021 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc
     4081 cgccgggatc actctcggca tggacgagct gtacaagtaa accggtgctc gaggtaccga
     4141 tgaatagcta aggtagaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga
     4201 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     4261 tggtcttact gacatccact ttgcctttct ctccacaggt gtcgacaatc aacctctgga
     4321 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4381 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4441 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4501 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4561 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4621 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4681 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4741 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4801 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4861 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc ggtacgatca
     4921 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4981 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     5041 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     5101 gaggattggg aagacaatag cccagctttt gttcccttta gtgagggtta attgcgcgct
     5161 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgcta attcactcct
     5221 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     5281 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     5341 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     5401 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     5461 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     5521 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     5581 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     5641 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     5701 ctcccagtca tagctgtccc tcttctctta tggagatcgg ccggccgccg gcgtcgacgc
     5761 gcgcggccgc ggcgcgccgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6421 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6481 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6541 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6601 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6661 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6721 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6781 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6841 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6901 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6961 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7021 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7081 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7141 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7201 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7261 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7321 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7381 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7441 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7501 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7561 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tacgcgcgcg
     7621 ccggcggcgc gccgtcgacg gccggccgcg gccgc
//
