LOCUS       sequence-252311-        8284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N2H assay vector with nanoluc fragment1 (empty control, no Gateway 
            cloning site, expressing nanoluc fragment1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8284)
  AUTHORS   Choi SG, Olivet J, Cassonnet P, Vidalain PO, Luck K, Lambourne L, 
            Spirohn K, Lemmens I, Dos Santos M, Demeret C, Jones L, Rangarajan 
            S, Bian W, Coutant EP, Janin YL, van der Werf S, Trepte P, Wanker 
            EE, De Las Rivas J, Tavernier J, Twizere JC, Hao T, Hill DE, Vidal 
            M, Calderwood MA, Jacob Y
  TITLE     Maximizing binary interactome mapping with a minimal number of 
            assays.
  JOURNAL   Nat Commun. 2019 Aug 29;10(1):3907. doi: 10.1038/s41467-019-11809-2.
  PUBMED    31467278
REFERENCE   2  (bases 1 to 8284)
  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"; journalName: "Nat 
            Commun."; date: "2019-08-29"; volume: "10(1):3907. doi"; pages: " 
            10.1038/s41467-019-11809-2"
COMMENT     Sequence Label: pDEST-N2H-F1-empty vector
FEATURES             Location/Qualifiers
     source          1..8284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     757..776
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        757..775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1186..1219)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1234..1355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1245..1264)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1299..1318
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1420..1607
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1475..1499)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(1612..1645)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     rep_origin      1660..1795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1894..1911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2065..2653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2145..2164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2824..3615,3616..3684))
                     /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     3447..3466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3685..3789)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3816..5158
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4779..4826)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     5199..5217
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5255..5277)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5377..5396
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5890..6984)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(6985..7392)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(7693..8148)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7830..7851)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8042..8061
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgg gcgcgcccct
      541 ccttgaatta attaaggcgc gccactgaaa tttttaatta aggcgcgcct agcatgtgat
      601 taattaagca tgtctttaga tctatgatga cacaaacccc gcccagcgtc ttgtcattgg
      661 cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatataa
      721 aaggtgacgc gtgtggatcc tcgaacaccg agctcttaat acgactcact atagggaaag
      781 caaaaatgtg atcttgcttg taaatacaat tttgagaggt taataaatta caagtagtgc
      841 tatttttgta tttaggttag ctatttagct ttacgttcca ggatgcctag tggcagcccc
      901 acaatatcca ggaagccctc tctgcggttt ttcagattag gtagtcgaaa aacctaagaa
      961 atttacctat gggcgtcttc acactcgaag atttcgttgg ggactggcga cagacagccg
     1021 gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg
     1081 gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg
     1141 acatccatgt catcatcccg tatgaaggtt aataagcatg cgaacataac ttcgtatagc
     1201 atacattata cgaagttatg gaagaagaca tgataagata cattgatgag tttggacaaa
     1261 ccacaacaag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1321 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1381 tgtttcaggt tcagggggag atgtgggagg ttttttaaag cgaatttctt atgatttatg
     1441 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
     1501 taggttttaa aacgaaaatt cttgttcttg agtaactctt tcctgtaggt caggttgctt
     1561 tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcgg aataacttcg
     1621 tatagtatac cttatacgaa gttatgccta actccgccca tcccgcccct aactccgccc
     1681 agttccgccc attctccgcc tcatggctga ctaatttttt ttatttatgc agaggccgag
     1741 gccgcctcgg cctctgagct attccagaag tagtgaggac gcttttttgg aggccgaggc
     1801 ttttgcaaag atcgaacaag agacaggact gcattaatga atcggccaac gcgcggggag
     1861 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1921 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1981 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2041 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2101 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2161 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2221 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2281 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2341 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2401 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2461 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2521 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2581 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2641 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2701 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2761 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2821 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2881 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2941 ccccagtgct gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat
     3001 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3061 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3121 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3181 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3241 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3301 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3361 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3421 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3481 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3541 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3601 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3661 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3721 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3781 ggttccgcgc acatttcccc gaaaagtgcc acctgaacga agcatctgtg cttcattttg
     3841 tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     3901 ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat
     3961 ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     4021 catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac
     4081 ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta
     4141 gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact
     4201 gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa
     4261 agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca
     4321 agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca
     4381 gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt
     4441 ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc
     4501 tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa
     4561 aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat
     4621 atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag
     4681 cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag
     4741 tgcgtcttca gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag
     4801 aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg
     4861 caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc
     4921 tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt
     4981 atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat
     5041 tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc
     5101 actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat
     5161 actaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5221 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5281 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5341 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5401 gagtgcacca tatcgactac gtcgttaagg ccgtttctga cagagtaaaa ttcttgaggg
     5461 aactttcacc attatgggaa atggttcaag aaggtattga cttaaactcc atcaaatggt
     5521 caggtcattg agtgtttttt atttgttgta tttttttttt tttagagaaa atcctccaat
     5581 atataaatta ggaatcatag tttcatgatt ttctgttaca cctaactttt tgtgtggtgc
     5641 cctcctcctt gtcaatatta atgttaaagt gcaattcttt ttccttatca cgttgagcca
     5701 ttagtatcaa tttgcttacc tgtattcctt tacatcctcc tttttctcct tcttgataaa
     5761 tgtatgtaga ttgcgtatat agtttcgtct accctatgaa catattccat tttgtaattt
     5821 cgtgtcgttt ctattatgaa tttcatttat aaagtttatg tacaaatatc ataaaaaaag
     5881 agaatctttt taagcaagga ttttcttaac ttcttcggcg acagcatcac cgacttcggt
     5941 ggtactgttg gaaccaccta aatcaccagt tctgatacct gcatccaaaa cctttttaac
     6001 tgcatcttca atggccttac cttcttcagg caagttcaat gacaatttca acatcattgc
     6061 agcagacaag atagtggcga tagggttgac cttattcttt ggcaaatctg gagcagaacc
     6121 gtggcatggt tcgtacaaac caaatgcggt gttcttgtct ggcaaagagg ccaaggacgc
     6181 agatggcaac aaacccaagg aacctgggat aacggaggct tcatcggaga tgatatcacc
     6241 aaacatgttg ctggtgatta taataccatt taggtgggtt gggttcttaa ctaggatcat
     6301 ggcggcagaa tcaatcaatt gatgttgaac cttcaatgta gggaattcgt tcttgatggt
     6361 ttcctccaca gtttttctcc ataatcttga agaggccaaa acattagctt tatccaagga
     6421 ccaaataggc aatggtggct catgttgtag ggccatgaaa gcggccattc ttgtgattct
     6481 ttgcacttct ggaacggtgt attgttcact atcccaagcg acaccatcac catcgtcttc
     6541 ctttctctta ccaaagtaaa tacctcccac taattctctg acaacaacga agtcagtacc
     6601 tttagcaaat tgtggcttga ttggagataa gtctaaaaga gagtcggatg caaagttaca
     6661 tggtcttaag ttggcgtaca attgaagttc tttacggatt tttagtaaac cttgttcagg
     6721 tctaacacta ccggtacccc atttaggacc acccacagca cctaacaaaa cggcatcagc
     6781 cttcttggag gcttccagcg cctcatctgg aagtggaaca cctgtagcat cgatagcagc
     6841 accaccaatt aaatgatttt cgaaatcgaa cttgacattg gaacgaacat cagaaatagc
     6901 tttaagaacc ttaatggctt cggctgtgat ttcttgacca acgtggtcac ctggcaaaac
     6961 gacgatcttc ttaggggcag acattagaat ggtatatcct tgaaatatat atatatatat
     7021 tgctgaaatg taaaaggtaa gaaaagttag aaagtaagac gattgctaac cacctattgg
     7081 aaaaaacaat aggtccttaa ataatattgt caacttcaag tattgtgatg caagcattta
     7141 gtcatgaacg cttctctatt ctatatgaaa agccggttcc ggcgctctca cctttccttt
     7201 ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg cgacctctga aattaacaaa
     7261 aaatttccag tcatcgaatt tgattctgtg cgatagcgcc cctgtgtgtt ctcgttatgt
     7321 tgaggaaaaa aataatggtt gctaagagat tcgaactctt gcatcttacg atacctgagt
     7381 attcccacag ttaactgcgg tcaagatatt tcttgaatca ggcgccttag accgctcggc
     7441 caaacaacca attacttgtt gagaaataga gtataattat cctataaata taacgttttt
     7501 gaacacacat gaacaaggaa gtacaggaca attgattttg aagagaatgt ggattttgat
     7561 gtaattgttg ggattccatt tttaataagg caataatatt aggtatgtag atatactaga
     7621 agttctcctc gaccggtcga tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     7681 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     7741 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     7801 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     7861 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     7921 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     7981 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     8041 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     8101 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca atattggcca
     8161 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     8221 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     8281 tgtt
//
