LOCUS       sequence-317771-        4454 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 4454)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4454)
  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-379    FLAG UnaG
FEATURES             Location/Qualifiers
     source          1..4454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        173..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             756..779
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             786..1199
                     /codon_start=1
                     /product="bilirubin-binding green fluorescent protein from 
                     Anguilla japonica (Kumagai et al., 2013)"
                     /label=UnaG
                     /translation="VEKFVGTWKIADSHNFGEYLKAIGAPKELSDGGDATTPTLYISQK
                     DGDKMTVKIENGPPTFLDTQVKFKLGEEFDEFPSDRRKGVKSVVNLVGEKLVYVQKWDG
                     KETTYVREIKDGKLVVTLTMGDVVAVRSYRRATE"
     CDS             1206..1229
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1354..1475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1391..1410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1445..1464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1482..1937)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1619..1640)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1831..1850
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1964..2068
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2070..2427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2070..2261
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2093..2113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2278..2413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2462..3256
                     /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             2462..3256
                     /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(2516..2535)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3126..3145
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3444..3463)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3488..3535
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3864..4452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4353..4372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
       61 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
      721 agatctcgag ctcaagcttc gaattcgcca ccatggacta caaagacgat gacgataaag
      781 gaagtgtcga aaagttcgtg ggcacatgga agattgccga tagccacaac ttcggcgaat
      841 acctcaaagc tataggggca ccaaaagagc ttagcgatgg tggagatgcg acaactccca
      901 cactgtacat tagccagaaa gacggagaca agatgactgt caagatcgag aatgggcctc
      961 ccacctttct ggacacacag gttaagttca aactgggtga ggagtttgac gagtttccca
     1021 gtgatcgcag gaaaggggtg aagtctgtgg tgaatctggt aggcgagaaa ctcgtgtacg
     1081 tccagaagtg ggatgggaag gaaaccacgt atgtgcgaga gatcaaggac gggaagttgg
     1141 ttgtcaccct gaccatgggc gatgtggttg ctgtgcgctc atatcggaga gccactgaaa
     1201 ctggtgatta caaggatgac gatgacaagt aagcggccgc gactctagat cataatcagc
     1261 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1321 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1441 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     1501 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1561 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1621 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1681 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1741 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1801 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1861 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1921 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1981 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2041 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2101 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2161 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2221 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2281 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2341 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2401 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2461 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2521 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2581 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2641 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2701 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2761 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2821 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2881 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2941 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3001 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3061 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3121 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3181 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3241 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3301 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3361 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3421 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3481 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     3541 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3601 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3661 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     3721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3781 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3841 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3901 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3961 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4021 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4081 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4141 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4201 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4261 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4321 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4381 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4441 agcctatgga aaaa
//
