LOCUS       sequence-360902-        5606 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 5606)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5606)
  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-395    HA miRFP670[1] IRES miRFP670[2]
FEATURES             Location/Qualifiers
     source          1..5606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        165..468
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        469..672
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     623..643
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             748..774
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(1172..1746,1747..1749,1750..1758)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1339..1356)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1566..1585
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2347..2373
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2498..2619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2535..2554)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2589..2608
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2626..3081)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2763..2784)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2975..2994
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3108..3212
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3214..3571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3214..3405
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3237..3257)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3422..3557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3606..4400
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3606..4400
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3660..3679)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4270..4289
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4588..4607)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4632..4679
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5008..5596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5497..5516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
       61 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
      121 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      181 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      241 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      301 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      361 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      421 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      481 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      541 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      601 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      661 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg
      721 agctcaagct tcgaattcgc caccatgtat ccctatgatg ttccagacta tgctggaagt
      781 gtggctgggc atgctagcgg ttctcctgcc tttggcaccg ctagtcactc caactgcgag
      841 cacgaggaga tccatcttgc tgggagtatc cagcctcacg gagcccttct ggtcgttagc
      901 gaacacgatc accgtgtgat tcaggcaagc gcgaatgcag ccgagttcct caacctgggc
      961 agtgtgctgg gagttccctt ggccgaaatc gacggagatc tgctgatcaa gattctgccc
     1021 catctcgatc cgacagctga aggcatgcca gtagcagtta ggtgtcgcat tgggaatccg
     1081 tccaccgagt attgcggtct gatgcatagg ccaccagaag gagggctgat tatcgagctg
     1141 gaaagagcgg gaccctaaga attcagatcc gcccctctcc ctcccccccc cctaacgtta
     1201 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     1261 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     1321 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     1381 aagcagttcc tctggtagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     1441 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     1501 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     1561 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     1621 attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt tagtcgaggt
     1681 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     1741 gataatatgg ccacaaccat gagagcggga cccagtatcg acctgtctgg gacactcgca
     1801 ccagcactgg agcgcataag aactgctgga tcactgaggg ccctctgtga cgacacagtc
     1861 ttgctctttc agcaatgtac gggctatgat cgagtgatgg tgtaccgatt cgacgaacag
     1921 ggtcatggtc tggtcttttc cgagtgtcac gtgcctggcc tggagagcta ctttggcaac
     1981 cgctatccct cttctaccgt accccagatg gccaggcaac tttacgtccg gcaacgggtc
     2041 agagtgctgg tggatgtgac ctaccagcct gtgccactcg aacctcggct gagcccactc
     2101 actggacggg atctggacat gagcggctgc ttcctgaggt ccatgtcacc ttgccatctg
     2161 cagttcttga aggacatggg cgttcgagcc acactcgcag tctcactggt tgtaggcggg
     2221 aaactgtggg gtcttgtggt gtgtcaccac taccttccca ggttcatccg gttcgagttg
     2281 agagccatct gcaaacgttt ggccgagaga atagccaccc gcattactgc tctggaatcc
     2341 actggttacc cttatgacgt gcctgattat gcctaagcgg ccgcgactct agatcataat
     2401 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2461 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2521 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2581 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     2641 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2701 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2761 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2821 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2881 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2941 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3001 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3061 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3121 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3181 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3241 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3301 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3481 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3541 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3601 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3661 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3721 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3781 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3841 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3901 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3961 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4021 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4081 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4141 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4201 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4261 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4321 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4381 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4441 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4501 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4561 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4621 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4681 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4741 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4801 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4861 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4921 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4981 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5041 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5101 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5161 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5221 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5281 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5341 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5401 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5461 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5521 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5581 gcggagccta tggaaaaacg ccagca
//
