LOCUS       sequence-249532-        5417 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 5417)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5417)
  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-406 FLAG IFP1.4[1] Stuffer/InteinC IFP1....
FEATURES             Location/Qualifiers
     source          1..5417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             644..667
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2057..2080
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2205..2326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2242..2261)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2296..2315
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2333..2788)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2470..2491)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2682..2701
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2815..2919
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2921..3278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2921..3112
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2944..2964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3129..3264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3313..4107
                     /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             3313..4107
                     /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(3367..3386)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3977..3996
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4295..4314)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4339..4386
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4715..5303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5204..5223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgccacc atggactaca aagacgatga
      661 cgataaagga agtgctcggg atccattgcc tttcttccca cccttgtacc tcggtggacc
      721 agagatcacc accgagaact gcgagaggga accgatacac attcccgggt ccattcagcc
      781 ccatggtgcg ctgctgactg ctgacggaca ttcaggggaa gtgctgcaag tcagcctgaa
      841 tgcagcaacc tttctgggcc aagaacccac tgtgcttcgc gggcagacct tggctgctct
      901 gctgcctgag cagtggccag ccctccaagc agctcttcca cccggttgtc cagacgccct
      961 ccagtatcgt gccactctgg actggcctgc tgccggccac cttagcctga cagtgcacag
     1021 ggtagctgag ctgctcatcc tcgaattcga gcccaccgag gcatggtctt gtctgtccta
     1081 tgagactgag atcctgacag ttgagtatgg tctgcttccc ataggcaaga tcgtggagaa
     1141 aaggatcgaa tgtaccgtgt actcagtcga caataacggc aacatctaca ctcaacctgt
     1201 tgcacagtgg catgatagag gagaacagga agtgttcgag tactgcttgg aggatgggag
     1261 tctgatacga gccacgaaag accacaagtt catgaccgta gatggacaga tgctgccaat
     1321 tgacgagatc tttgagcggg aattggacct gatgagggtg gataatcttc cgaacatcaa
     1381 gattgctaca cgcaagtatc tcgggaaaca gaacgtctac gacattggtg tggaacgtga
     1441 ccacaacttt gcgctcaaga atggcttcat tgccagcaat tgcgacagca ttggacccca
     1501 tgcgctgagg aacgctatgt tcgcccttga atctgctccc aatctccgag cactggctga
     1561 agtcgccaca cagacagtca gggagctgac aggctttgat cgggtgatgc tgtacaagtt
     1621 tgccccagat gcaacgggtg agatgatcgc agaagccaga cgggaaggga tgcaggcctt
     1681 cctgggccac agatttccgg cctcacatac gcctgctcag gccagagcac tgtacacccg
     1741 acacctcctg agactgactg cggacactag agcagctgcc gtgccactcg atcccgtgct
     1801 caacccacag accaatgcgc ctaccccttt gggaggagcc gtactgaggg ccacatcccc
     1861 tatgcacatg cagtatctgc gcaacatggg cgttgggagt agtctgtctg tgtccgttgt
     1921 cgttggaggg caactgtggg gcttgatcgt ctgccaccat cagacaccgt atgtgcttcc
     1981 tcctgacttg cgtacaactc tggaggagct gggccgcaag cttagcggcc aggtgcagcg
     2041 gaaagaggcc actggtgatt acaaggatga cgatgacaag taagcggccg cgactctaga
     2101 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2161 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2221 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2281 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     2341 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2401 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2461 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2521 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     2581 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2641 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2701 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2761 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2821 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2881 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     2941 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3001 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3061 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3121 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3181 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3241 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3301 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3361 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3421 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3481 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3541 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3601 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3661 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3721 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3781 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3841 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3901 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3961 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4021 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4081 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4141 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4201 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4261 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4321 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4381 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4441 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     4501 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     4561 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4621 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4681 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4741 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4801 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4861 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4921 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4981 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5041 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5101 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5161 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5221 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5281 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5341 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5401 gtattaccgc catgcat
//
