LOCUS       sequence-249531-        5171 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 5171)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5171)
  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-405 FLAG Venus210[1] Stuffer/InteinC
            Ven...
FEATURES             Location/Qualifiers
     source          1..5171
                     /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"
     primer_bind     complement(977..996)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1811..1834
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1959..2080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1996..2015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2050..2069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2087..2542)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2224..2245)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2436..2455
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2569..2673
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2675..3032
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2675..2866
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2698..2718)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2883..3018
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3067..3861
                     /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             3067..3861
                     /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(3121..3140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3731..3750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4049..4068)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4093..4140
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4469..5057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4958..4977
                     /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 agtgtgtcaa agggagaaga gttgtttacc ggtgttgttc ccatactggt
      721 ggaactggat ggtgacgtca atggacacaa gtttagcgtt tctggcgaag gagaaggcga
      781 tgctacctat ggcaaactga ccctcaaact gatctgtacg actggaaagt tgcctgtacc
      841 ttggccaaca ctggtcacaa ccttgggcta tgggctgcaa tgctttgccc gttatcctga
      901 tcacatgaag caacatgact tctttaagtc tgctatgcca gagggatatg tgcaggaacg
      961 gactatcttc ttcaaggacg acggcaacta caagacgaga gctgaggtga aatttgaggg
     1021 tgacacactg gtgaatcgca tagagctgaa aggcatcgac ttcaaagaag atgggaatat
     1081 tctcgggcat aaactcgagt acaactacaa ttcccacaac gtgtacatca ctgccgataa
     1141 gcagaagaat gggattaagg cgaacttcaa gatccgacac aacatcgagg atggtggggt
     1201 ccagcttgcc gaccattatc agcagaacac acccattggg gatggaccag tcctgttgcc
     1261 ggataaccac tacctcagtt accagagcaa actgtccaaa tcttgtctgt cctatgagac
     1321 tgagatcctg acagttgagt atggtctgct tcccataggc aagatcgtgg agaaaaggat
     1381 cgaatgtacc gtgtactcag tcgacaataa cggcaacatc tacactcaac ctgttgcaca
     1441 gtggcatgat agaggagaac aggaagtgtt cgagtactgc ttggaggatg ggagtctgat
     1501 acgagccacg aaagaccaca agttcatgac cgtagatgga cagatgctgc caattgacga
     1561 gatctttgag cgggaattgg acctgatgag ggtggataat cttccgaaca tcaagattgc
     1621 tacacgcaag tatctcggga aacagaacgt ctacgacatt ggtgtggaac gtgaccacaa
     1681 ctttgcgctc aagaatggct tcattgccag caattgcgat cccaatgaga agagggacca
     1741 tatggtactg ctcgaattcg tgactgcagc aggcattacc cttgggatgg atgaacttta
     1801 taagactggt gattacaagg atgacgatga caagtaagcg gccgcgactc tagatcataa
     1861 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1921 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1981 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2041 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2101 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2161 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2221 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2281 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2341 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2401 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2461 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2521 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2581 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2641 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2701 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2761 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2821 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2881 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2941 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3001 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3061 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3121 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3181 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3241 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3301 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3361 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3421 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3481 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3541 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3601 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3661 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3721 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3781 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3841 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3901 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3961 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4021 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4081 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4141 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4201 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4261 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4321 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4381 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4441 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4501 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4561 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4621 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4681 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4741 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4801 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4861 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4921 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4981 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5041 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5101 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5161 ccgccatgca t
//
