LOCUS       sequence-252313-        7230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N2H assay vector with nanoluc fragment2 (empty control, no Gateway 
            cloning site, expressing nanoluc fragment2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7230)
  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 7230)
  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-F2-empty vector
FEATURES             Location/Qualifiers
     source          1..7230
                     /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    1306..1339
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1387..1508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1398..1417)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1452..1471
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1573..1760
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1628..1652)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      1777..1912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2011..2028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2182..2770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2262..2281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2941..3732,3733..3801))
                     /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     3564..3583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3802..3906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3933..5275
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4896..4943)
                     /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)."
     protein_bind    5298..5331
                     /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."
     primer_bind     5362..5380
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5418..5440)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5540..5559
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5580..5861
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5862..6536
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(6639..7094)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6776..6797)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6988..7007
                     /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 gggcctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg
     1021 tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg
     1081 acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt
     1141 tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg
     1201 agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg
     1261 gctggcggct gtgcgaacgc attctggcgt aataagcatg cgaacataac ttcgtataat
     1321 gtatgctata cgaagttatg gaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaagagaag
     1381 acatgataag atacattgat gagtttggac aaaccacaac aagaatgcag tgaaaaaaat
     1441 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1501 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     1561 aggtttttta aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     1621 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttgttc
     1681 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt
     1741 attgaccaca cctctaccgg cggcctaact ccgcccatcc cgcccctaac tccgcccagt
     1801 tccgcccatt ctccgcctca tggctgacta atttttttta tttatgcaga ggccgaggcc
     1861 gcctcggcct ctgagctatt ccagaagtag tgaggacgct tttttggagg ccgaggcttt
     1921 tgcaaagatc gaacaagaga caggactgca ttaatgaatc ggccaacgcg cggggagagg
     1981 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2041 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2101 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2161 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2221 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2281 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2341 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2401 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2461 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2521 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2581 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2641 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2701 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2761 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2821 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2881 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2941 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3001 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3061 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3121 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3181 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3241 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3301 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3361 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3421 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3481 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3541 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3601 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3661 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3721 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3781 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3841 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3901 tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt cattttgtag
     3961 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4021 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     4081 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     4141 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     4201 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     4261 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     4321 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     4381 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     4441 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     4501 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     4561 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     4621 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     4681 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     4741 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     4801 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     4861 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     4921 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     4981 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     5041 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     5101 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     5161 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     5221 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatact
     5281 aagaaaccat tggtaccata acttcgtata aggtatacta tacgaagtta tgctagcatt
     5341 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5401 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5461 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5521 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagatca
     5581 acgacattac tatatatata atataggaag catttaatag aacagcatcg taatatatgt
     5641 gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag
     5701 cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta
     5761 attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca
     5821 cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca
     5881 caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat
     5941 gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa
     6001 caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt
     6061 caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg
     6121 ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag
     6181 aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca
     6241 acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag
     6301 aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc
     6361 aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg
     6421 gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa
     6481 atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta
     6541 ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa
     6601 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     6661 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6721 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6781 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6841 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6901 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6961 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     7021 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     7081 gctacagggc gcgtcaatat tggccattag ccatattatt cattggttat atagcataaa
     7141 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     7201 ttggctcatg tccaatatga ccgccatgtt
//
