LOCUS       sequence-249205-        6573 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 6573)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6573)
  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-324    N-Tag IFP1.4[2] L32 {Esp3I(2x)} EGFP
FEATURES             Location/Qualifiers
     source          1..6573
                     /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(1732..2306,2307..2309,2310..2318)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1899..1916)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2126..2145
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2319..2321,2322..2324,2325..3038)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2319..2321,2322..2324,2325..3035)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2364..2385)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2625..2644)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2972..2993
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3084..3216
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3225..3813
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3278..3298)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3696..3707)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3840..3857)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3846..3957
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4053..4073)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4053..4071)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4092..4108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4092..4108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4105..4127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4210..4229)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4275..4330
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4329..4348)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4709..5297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4789..4808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5468..6259,6260..6328))
                     /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     6091..6110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6329..6433)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6501..6519
                     /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 caccatggac agcattggac cccatgcgct gaggaacgct atgttcgccc ttgaatctgc
     1081 tcccaatctc cgagcactgg ctgaagtcgc cacacagaca gtcagggagc tgacaggctt
     1141 tgatcgggtg atgctgtaca agtttgcccc agatgcaacg ggtgagatga tcgcagaagc
     1201 cagacgggaa gggatgcagg ccttcctggg ccacagattt ccggcctcac atacgcctgc
     1261 tcaggccaga gcactgtaca cccgacacct cctgagactg actgcggaca ctagagcagc
     1321 tgccgtgcca ctcgatcccg tgctcaaccc acagaccaat gcgcctaccc ctttgggagg
     1381 agccgtactg agggccacat cccctatgca catgcagtat ctgcgcaaca tgggcgttgg
     1441 gagtagtctg tctgtgtccg ttgtcgttgg agggcaactg tggggcttga tcgtctgcca
     1501 ccatcagaca ccgtatgtgc ttcctcctga cttgcgtaca actctggagg agctgggccg
     1561 caagcttagc ggccaggtgc agcggaaaga ggccgacgtg ggtggcggag ggagcgaagg
     1621 cggcggatcc ggagggcctg gctctggcgg agaggggagc gctggaggcg ggtcagccgg
     1681 tggtgggagt cgagacgccg aggcacgccg tctcctaaga attcagatcc gcccctctcc
     1741 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     1801 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     1861 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     1921 tctgttgaat gtcgtgaagg aagcagttcc tctggtagct tcttgaagac aaacaacgtc
     1981 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     2041 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     2101 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     2161 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt
     2221 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     2281 tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc
     2341 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2401 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2461 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2521 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2581 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2641 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2701 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2761 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2821 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2881 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     2941 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3001 ccgggatcac tctcggcatg gacgagctgt acaagtaaac cggtgctcga ggtaccgatg
     3061 aatagctaag gtcgaggccg caggtaagta tcaaggttac aagacaggtt taaggagacc
     3121 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     3181 gtcttactga catccacttt gcctttctct ccacaggtgt cgacaatcaa cctctggatt
     3241 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3301 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3361 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3421 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3481 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3541 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3601 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3661 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3721 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3781 cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacgatcagc
     3841 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3901 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3961 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4021 ggattgggaa gacaatagcc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     4081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctaat tcactcctca
     4141 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     4201 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     4261 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4321 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     4381 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     4441 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     4501 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     4561 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     4621 cccagtcata gctgtccctc ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc
     4681 gcggccgcgg cgcgccgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4741 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4801 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4861 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4921 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4981 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5041 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5101 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5161 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5221 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5281 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5341 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5401 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5461 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5521 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5581 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5641 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5701 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5761 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5821 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5881 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5941 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6001 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6061 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6121 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6181 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6241 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6301 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6481 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc
     6541 ggcggcgcgc cgtcgacggc cggccgcggc cgc
//
