LOCUS       sequence-367187-       10522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length CRM1 fused to binding domain of two-hybrid plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10522)
  AUTHORS   Belanger KD, Yewdell WT, Barber MF, Russo AN, Pettit MA, Damuth EK, 
            Hussain N, Geier SJ, Belanger KG
  TITLE     Exportin Crm1 is important for Swi6 nuclear shuttling and MBF 
            transcription activation in Saccharomyces cerevisiae.
  JOURNAL   BMC Mol Cell Biol. 2022 Feb 21;23(1):10. doi: 
            10.1186/s12860-022-00409-6.
  PUBMED    35189816
REFERENCE   2  (bases 1 to 10522)
  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: "BMC Mol 
            Cell Biol."; date: "2022-02-21"; volume: "23(1):10. doi"; pages: " 
            10.1186/s12860-022-00409-6"
COMMENT     Sequence Label: KBB486
FEATURES             Location/Qualifiers
     source          1..10522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        803..1507
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1535..1975
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     terminator      5712..5899
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5767..5791)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    6729..7891
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      7054..7891
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(7730..8404)
                     /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"
     promoter        complement(8405..8506)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8526..8544)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8612..8716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8717..8785,8786..9577)
                     /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     complement(8935..8954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9748..10336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10237..10256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10490..10507
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag gatccgggat
       61 cgaagaaatg atggtaaatg aaataggaaa tcaaggagca tgaaggcaaa agacaaatat
      121 aagggtcgaa cgaaaaataa agtgaaaagt gttgatatga tgtatttggc tttgcggcgc
      181 cgaaaaaacg agtttacgca attgcacaat catgctgact ctgtggcgga cccgcgctct
      241 tgccggcccg gcgataacgc tgggcgtgag gctgtgcccg gcggagtttt ttgcgcctgc
      301 attttccaag gtttaccctg cgctaagggg cgagattgga gaagcaataa gaatgccggt
      361 tggggttgcg atgatgacga ccacgacaac tggtgtcatt atttaagttg ccgaaagaac
      421 ctgagtgcat ttgcaacatg agtatactag aagaatgagc caagacttgc gagacgcgag
      481 tttgccggtg gtgcgaacaa tagagcgacc atgaccttga aggtgagacg cgcataaccg
      541 ctagagtact ttgaagagga aacagcaata gggttgctac cagtataaat agacaggtac
      601 atacaacact ggaaatggtt gtctgtttga gtacgctttc aattcatttg ggtgtgcact
      661 ttattatgtt acaatatgga agggaacttt acacttctcc tatgcacata tattaattaa
      721 agtccaatgc tagtagagaa ggggggtaac acccctccgc gctcttttcc gatttttttc
      781 taaaccgtgg aatatttcgg atatcctttt gttgtttccg ggtgtacaat atggacttcc
      841 tcttttctgg caaccaaacc catacatcgg gattcctata ataccttcgt tggtctccct
      901 aacatgtagg tggcggaggg gagatataca atagaacaga taccagacaa gacataatgg
      961 gctaaacaag actacaccaa ttacactgcc tcattgatgg tggtacataa cgaactaata
     1021 ctgtagccct agacttgata gccatcatca tatcgaagtt tcactaccct ttttccattt
     1081 gccatctatt gaagtaataa taggcgcatg caacttcttt tctttttttt tcttttctct
     1141 ctcccccgtt gttgtctcac catatccgca atgacaaaaa aatgatggaa gacactaaag
     1201 gaaaaaatta acgacaaaga cagcaccaac agatgtcgtt gttccagagc tgatgagggg
     1261 tatctcgaag cacacgaaac tttttccttc cttcattcac gcacactact ctctaatgag
     1321 caacggtata cggccttcct tccagttact tgaatttgaa ataaaaaaaa gtttgctgtc
     1381 ttgctatcaa gtataaatag acctgcaatt attaatcttt tgtttcctcg tcattgttct
     1441 cgttcccttt cttccttgtt tctttttctg cacaatattt caagctatac caagcataca
     1501 atcaactcca agcttgaagc aagcctcctg aaagatgaag ctactgtctt ctatcgaaca
     1561 agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc
     1621 caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct
     1681 gactagggca catctgacag aagtggaatc aaggctagaa agactggaac agctatttct
     1741 actgattttt cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat
     1801 aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga
     1861 tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc
     1921 gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcgaacta
     1981 tctattcgat gatgaagata ccccaccaaa cccaaaaaaa gagatcgaat tccagctgac
     2041 caccatggaa ggaattttgg atttttctaa cgacttagat atcgctctat tagatcaggt
     2101 ggtatctaca ttctatcaag gttcaggtgt tcagcaaaag caagcccaag agattttgac
     2161 taaatttcaa gataatccag acgcttggca aaaagctgac caaattcttc aattctcaac
     2221 taaccctcaa tcaaaattta ttgccctttc catccttgat aaattaatta ctagaaaatg
     2281 gaaattgttg ccgaatgatc atagaattgg aattaggaac ttcgttgttg gtatgattat
     2341 ctccatgtgc caagacgatg aagtatttaa aacacaaaag aacctgatca ataagtcgga
     2401 cttgactttg gtccaaattt taaagcagga atggcctcag aattggcctg aatttatccc
     2461 agaactaatc ggcagttcaa gttcctctgt taatgtttgt gaaaataaca tgatcgtttt
     2521 gaaactgcta tccgaagaag tcttcgattt ctctgcggaa caaatgacac aagctaaagc
     2581 tttacatttg aaaaattcca tgtcaaaaga atttgaacag atttttaaat tatgttttca
     2641 agttttggaa caaggttctt caagctctct aatagtggca accttagaat ctttactgag
     2701 atatttacat tggattcctt atcgttatat ttatgaaacc aatattttgg aattattaag
     2761 caccaaattc atgacatcgc ctgatacaag agccatcaca ttgaaatgtt tgactgaggt
     2821 ttcaaatcta aaaattccac aagataatga tttgataaaa agacaaactg tacttttttt
     2881 ccaaaatact ctacaacaaa ttgccacaag tgtgatgccc gtgactgccg atctgaaagc
     2941 cacttatgca aatgctaacg gtaatgatca atctttctta caagatttag caatgtttct
     3001 aactacatat ctcgctcgta atagggccct tctagagagt gatgaatcgt taagagagtt
     3061 gttgttgaat gcgcaccaat acttaattca attatccaaa atcgaagaaa gagaactatt
     3121 taagacgaca ttggattact ggcataattt agtagcagat ttgttttatg aggttcagcg
     3181 tttgcctgct accgaaatga gcccattgat acagttatca gtcggttcac aagctatctc
     3241 aactggatct ggcgccctaa atccggaata tatgaaaaga tttccattaa aaaaacatat
     3301 ttacgaagaa atttgttcac agttgagatt ggtcattatt gaaaatatgg ttagaccaga
     3361 agaagtcctt gtggttgaaa atgatgaagg agaaattgtt agagagttcg ttaaagaatc
     3421 agacactatc caattataca aatcagaaag agaggttctt gtatatttga cccatttgaa
     3481 tgttatcgat acagaagaaa tcatgatcag taaattggca aggcaaatcg atggttccga
     3541 atggtcatgg cataatatca acactttatc ttgggctatt ggttccatat ctggtacaat
     3601 gagcgaagat actgaaaaaa ggtttgtagt tactgttatt aaagatttac tagatttaac
     3661 ggtcaagaaa aggggtaaag ataataaagc tgttgttgca tcggatatta tgtacgtcgt
     3721 gggccaatat cctagatttt tgaaggctca ctggaatttt ctaagaacag ttattttgaa
     3781 actttttgaa tttatgcatg aaactcatga gggtgttcag gatatggcat gtgacacatt
     3841 cattaagatt gttcaaaaat gtaaatatca ttttgttatt caacaacccc gtgagtctga
     3901 gcctttcatc caaacaataa ttagggatat tcaaaagaca actgctgacc tacagccgca
     3961 acaggtccat acattttaca aggcttgtgg tattattatt tctgaagaaa gaagcgtagc
     4021 ggagagaaat agattattaa gtgatttgat gcaactgcca aatatggctt gggacactat
     4081 agtggaacaa tccactgcta acccaacatt gttgttagat tctgaaactg tcaaaattat
     4141 tgcgaatatt ataaaaacca atgtagctgt ctgcacctca atgggagcag atttctatcc
     4201 acagttgggt cacatctatt acaatatgct tcagctatac agagctgttt cttcgatgat
     4261 ttctgctcaa gttgccgcag aaggtctaat tgctacgaag acaccaaagg ttcgtggttt
     4321 gagaaccatc aaaaaagaga ttctgaagct tgttgagacc tatatatcca aggcgagaaa
     4381 tttggatgat gtcgttaaag tgttagttga gccattattg aacgctgtac ttgaagacta
     4441 tatgaacaac gttcctgatg caagggacgc cgaagtgcta aattgtatga caacagtagt
     4501 agaaaaggtt ggtcatatga tcccacaagg tgttatttta attttacaga gtgtatttga
     4561 atgcactttg gacatgatta acaaagactt cacagaatat ccagagcatc gtgtagaatt
     4621 ttacaagtta ttgaaggtca tcaatgagaa atcatttgct gcatttttgg aactacctcc
     4681 ggctgccttt aaattgtttg ttgacgctat ttgttgggcg ttcaaacata acaatagaga
     4741 tgtcgaagtt aatggtctac agattgcatt agacctagtg aaaaacattg aaagaatggg
     4801 aaatgtccca tttgcaaatg agttccacaa gaattatttt ttcatttttg tcagtgaaac
     4861 gttttttgtt ttaaccgatt ccgaccacaa atccggtttt tctaagcaag ctttgttact
     4921 aatgaagttg atttctttgg tttatgataa caagatttcg gtaccattat atcaagaagc
     4981 tgaggtacca caaggaactt caaatcaagt ctacttaagc caatatttgg ctaatatgtt
     5041 aagtaatgca ttcccccatt taacatccga acaaatcgca agttttctct ctgcattaac
     5101 taaacaatac aaagacttag tggtcttcaa aggcactttg agggactttt tggtgcaaat
     5161 caaagaagtc ggaggtgatc caacagatta tttgtttgct gaagataaag agaatgcatt
     5221 aatggaacag aatagactag agagggaaaa agctgccaag attggtgggt tattaaaacc
     5281 ttccgaactt gatgattaca tggcaattcc cggggatccg tcgacctgca gagatctatg
     5341 aatcgtagat actgaaaaac cccgcaagtt cacttcaact gtgcatcgtg caccatctca
     5401 atttctttca tttatacatc gttttgcctt cttttatgta actatactcc tctaagtttc
     5461 aatcttggcc atgtaacctc tgatctatag aattttttaa atgactagaa ttaatgccca
     5521 tctttttttt ggacctaaat tcttcatgaa aatatattac gagggcttat tcagaagctt
     5581 tggacttctt cgccagaggt ttggtcaagt ctccaatcaa ggttgtcggc ttgtctacct
     5641 tgccagaaat ttacgaaaag atggaaaagg gtcaaatcgt tggtagatac gttgttgaca
     5701 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
     5761 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct
     5821 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagcatgag gtcgctctta
     5881 ttgaccacac ctctaccggc atgccgagca aatgcctgca aatcgctccc catttcaccc
     5941 aattgtagat atgctaactc cagcaatgag ttgatgaatc tcggtgtgta ttttatgtcc
     6001 tcagaggaca acacctgttg taatcgttct tccacacgga tcctggcgta atagcgaaga
     6061 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     6121 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataatcg ctgggccatt
     6181 ctcatgaaga atatcttgaa tttattgtca tattactagt tggtgtggaa gtcctaatat
     6241 cggtgatcaa tatagtggtt gacatgctgg ctagtcaaca ttgagccttt tgatcatgca
     6301 aatatattac ggtattttac aatcaaatat caaacttaac tattgacttt ataacttatt
     6361 taggtggtaa cattcttata aaaaagaaaa aaattactgc aaaacagtac tagcttttaa
     6421 cttgtatcct aggttatcta tgctgtctca ccatagagaa tattacctat ttcagaatgt
     6481 atgtccatga ttcgccgggt aaatacatat aatacacaaa tctggcttaa taaagtctat
     6541 aatatatctc ataaagaagt gctaaattgg ctagtgctat atatttttaa gaaaatttct
     6601 tttgactaag tccatatcga ctttgtaaaa gttcacatta gcatacatat attacacgag
     6661 ccagaaatag taacttttgc ctaaatcaca aattgcaaaa tttaattgct tgcaaaaggt
     6721 cacatgctta taatcaactt ttttaaaaat ttaaaatact tttttatttt ttatttttaa
     6781 acataaatga aataatttat ttattgttta tgattaccga aacataaaac ctgctcaaga
     6841 aaaagaaact gttttgtcct tggaaaaaaa gcactaccta ggagcggcca aaatgccgag
     6901 gctttcatag cttaaactct ttacagaaaa taggcattat agatcagttc gagttttctt
     6961 attcttcctt ccggttttat cgtcacagtt ttacagtaaa taagtatcac ctcttagagt
     7021 tcgatgataa gctgtcaaac atgagaatta attccacatg ttaaaatagt gaaggagcat
     7081 gttcggcaca cagtggaccg aacgtggggt aagtgcacta gggtccggtt aaacggatct
     7141 cgcattgatg aggcaacgct aattatcaac atatagattg ttatctatct gcatgaacac
     7201 gaaatcttta cttgacgact tgaggctgat ggtgtttatg caaagaaacc actgtgttta
     7261 atatgtgtca ctgtttgata ttactgtcag cgtagaagat aatagtaaaa gcggttaata
     7321 agtgtatttg agataagtgt gataaagttt ttacagcgaa aagacgataa atacaagaaa
     7381 atgattacga ggatacggag agaggtatgt acatgtgtat ttatatacta agctgccggc
     7441 ggttgtttgc aagaccgaga aaaggctagc aagaatcggg tcattgtagc gtatgcgcct
     7501 gtgaacattc tcttcaacaa gtttgattcc attgcggtga aatggtaaaa gtcaaccccc
     7561 tgcgatgtat attttcctgt acaatcaatc aaaaagccaa atgatttagc attatcttta
     7621 catcttgtta ttttacagat tttatgttta gatcttttat gcttgctttt caaaaggctt
     7681 gcaggcaagt gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag
     7741 catttttgac gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc
     7801 cgcttacatc aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg
     7861 tcagtccacc agctaacata aaatgtaagc tctcggggct ctcttgcctt ccaacccagt
     7921 cagaaatcga gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg
     7981 gaataaacga atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa
     8041 atacgagtct tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat
     8101 ctccatgcag ttggacgata tcaatgccgt aatcattgac cagagccaaa acatcctcct
     8161 taggttgatt acgaaacacg ccaaccaagt atttcggagt gcctgaacta tttttatatg
     8221 cttttacaag acttgaaatt ttccttgcaa taaccgggtc aattgttctc tttctattgg
     8281 gcacacatat aatacccagc aagtcagcat cggaatctag tgcacattct gcggcctctg
     8341 tgctctgcaa gccgcaaact ttcaccaatg gaccagaact acctgtgaaa ttaataacag
     8401 acatactcca agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc
     8461 aatgccctcc ctcttggccc tctccttttc ttttttcgac cgaattaatt cttgaagacg
     8521 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     8581 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     8641 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     8701 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     8761 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     8821 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     8881 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     8941 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     9001 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     9061 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     9121 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     9181 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     9241 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     9301 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     9361 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     9421 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     9481 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     9541 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     9601 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     9661 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     9721 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     9781 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     9841 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     9901 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     9961 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
    10021 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
    10081 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
    10141 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
    10201 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
    10261 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
    10321 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
    10381 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
    10441 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
    10501 gagcgaggaa gcggaagagc gc
//
