LOCUS       sequence-367189-        8689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SWI6-GFP full length under control of GAL1 promotion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8689)
  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 8689)
  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: KBB484
FEATURES             Location/Qualifiers
     source          1..8689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      695..1283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1184..1203
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1437..1454
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1571..1592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1607..1612,1613..1630,1631..1637)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1645..1661
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1650..1672
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1669..1685
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1669..1685
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1704..1724
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1706..1724
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2133..2156
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4592..5308
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGDILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(4625..4653)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5248..5268
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5371..5387)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5372..5388)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5390..5637
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5406..5424)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5655..5674)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5656..5674)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5684..5701)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5684..5700)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5693..5715)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5841..6296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5928..5947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6138..6159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6427..7230)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7231..7446)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7470..7489)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7589..7611
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7649..7667)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    7708..8211
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        8248..8352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8353..8421,8422..8689,1..524)
                     /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(8571..8590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
       61 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
      121 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
      181 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
      241 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
      301 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
      361 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
      421 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
      481 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
      541 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
      601 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
      661 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      721 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      781 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      841 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      901 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      961 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1021 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1081 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1141 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1201 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1261 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1321 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1381 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     1441 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     1501 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     1561 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     1621 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     1681 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     1741 tcgtgacagc cctccgaagg aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt
     1801 tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa caataaagat tctacaatac
     1861 tagcttttat ggttatgaag aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa
     1921 tgaacgaatc aaattaacaa ccataggatg ataatgcgat tagtttttta gccttatttc
     1981 tggggtaatt aatcagcgaa gcgatgattt ttgatctatt aacggatata taaatgcaaa
     2041 aactgcataa ccactttaac taatactttc aacattttcg gtttgtatta cttcttattc
     2101 aaatgtaata aaagtatcaa caaaaaattg ttaatatacc tctatacttt aacgtcaagg
     2161 agaaaaaact ataatggcgt tggaagaagt ggtacgatac ttaggacctc acaatgagat
     2221 ccctttgaca ctcactagag actcggagac tggccatttc ctcctgaaac attttctgcc
     2281 cattttgcag caatatcatg acacggggaa tattaacgag accaaccccg atagtttccc
     2341 cactgatgag gaaagaaata aattactggc acattatgga attgctgtaa atacagacga
     2401 ccgaggtgag ttatggatag agctggaaaa atgtttacaa ttattaaaca tgctaaattt
     2461 attcggtttg ttccaggatg cattcgaatt tgaggagcct gaaacagatc aggatgaaga
     2521 agatcccagc cattcgaaac taccagaaaa caagaccaaa agtgaaaatt ctaaggataa
     2581 tatcagctca aagaggatta ataatttaca agatatgagc ctggattctg atgcacacag
     2641 agaattaggc tctcctttaa aaaaactaaa aatagatacc tctgtaatag atgctgagag
     2701 tgactccact ccgaatactg ccagaggcaa gcctaacgat gatattaata agggccctag
     2761 cggcgacaat gaaagtaatg gcactgatga caatgacaga accgctggac ctatcataac
     2821 attcactcat gacctaactt ctgacttttt aagcagtcca ctgaaaatca tgaaagcact
     2881 accttctcca gttgtaaatg ataatgaaca gaagatgaaa ctagaggcat tcttacaacg
     2941 gttgttattt ccagaaattc aagaaatgcc tacatccctt aataatgaca gcagtaatag
     3001 aaattcagaa ggggggagct caaaccaaca acaacagcac gtatcatttg atagcctttt
     3061 gcaagaggta aacgacgctt ttcctaatac tcaattaaat cttaatattc ctgtagatga
     3121 gcatggaaac acaccattac attggctgac ttcaatagca aacctggaat tagtgaaaca
     3181 cctggttaag catggttcaa acagattata tggtgataat atgggggagt catgcctagt
     3241 gaaagctgtc aaatcagtaa ataattacga ctctggtact tttgaggcac ttctagatta
     3301 tttatatcca tgtttaattt tggaagactc aatgaataga acaattttgc accatatcat
     3361 tattacgtct ggtatgactg gctgttcagc agctgcgaaa tattatttag atattttaat
     3421 gggatggatt gtcaagaaac aaaatagacc cattcaaagt ggtactaacg aaaaagaaag
     3481 caaaccgaat gacaaaaatg gggaaagaaa ggactctata ctagaaaatt tggacctgaa
     3541 atggataata gcaaacatgc tcaatgcgca ggactctaag ggcgatactt gcttgaacat
     3601 tgcggcaaga ttgggaaaca tttcaatagt agacgcttta ttagattacg gtgctgaccc
     3661 atttattgca aacaaatcag ggttgagacc cgtggatttt ggggcaggta cttcaaaatt
     3721 acaaaatacg aacggcggtg acgaaaactc gaagatggtc tctaagggtg attacgacgg
     3781 tcaaaaaaat ggaaaggcca aaaaaataag gtctcaactg ttgaaaaacc cacctgaaac
     3841 aacttcgtta attaatgatg tccaaaattt actgaattca atctcgaaag attatgagaa
     3901 tgaaacagtg caatataatg agaaattaga aaaactacac aaggaattga acgaacagcg
     3961 agaagaatta gctaattcta gagaacaact agcaaatgtg aagcaattga aagatgaata
     4021 ctcactaatg caagagcaat tgaccaattt aaaagcagga attgaagaag aagaggaaag
     4081 ttttagggaa gagagcaaaa agctaggaat aattgcagat gaaagttcag gtattgattg
     4141 ggactctagt gaatatgatg cagatgaacc cttcaaagta gagttccttt cagatttctt
     4201 ggaggataaa ttacaaaaga attatgaagg cgatatttcc aaactattag aagcggagtc
     4261 gaaagagcag ataatggaac agatacgaaa tcaacttccc gcagaaaaaa tccaatcgat
     4321 gcttccacca acggtcttat tgaaggcaag gataaatgca tacaagcgga atgataaaca
     4381 cctaaccaat gtattggata caatcagcac caaacaatca gaattagaga ataaatttag
     4441 aagagtgtta tctttatgtt tgaaaatcga cgaaaataaa gttgacaata tgcttgatgg
     4501 tttgttacag gccatatctt ctgaagaccc acaagacatt gacactgacg aaatgcaaga
     4561 ttttttaaaa aagcatgctt caggtcgagc tatggctagc aaaggagaag aactcttcac
     4621 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
     4681 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
     4741 tactggaaaa ctacctgttc catggccaac acttgtcact actttcactt atggtgttca
     4801 gtgcttttca agatacccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc
     4861 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
     4921 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     4981 ttttaaagaa gatggagaca ttcttgggca caaattggaa tacaactata actcacacaa
     5041 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattagaca
     5101 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
     5161 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     5221 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     5281 tacacatggc atggatgaac tatacaaagg atccggctgc taaagaacta gtggatcccc
     5341 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagt catgtaatta
     5401 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     5461 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     5521 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     5581 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
     5641 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     5701 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5761 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5821 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5881 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5941 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6001 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6061 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6181 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6241 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     6301 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg
     6361 atataattaa attgaagctc taatttgtga gtttagtata catgcattta cttataatac
     6421 agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta
     6481 acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat
     6541 aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc
     6601 tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct
     6661 tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat
     6721 gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg acaattctgc
     6781 taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct
     6841 aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct
     6901 gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc
     6961 ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc
     7021 catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa
     7081 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
     7141 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
     7201 agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt
     7261 tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg
     7321 tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
     7381 cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
     7441 attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     7501 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     7561 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     7621 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     7681 gtcatgataa taatggtttc ttagacggat cgcttgcctg taacttacac gcgcctcgta
     7741 tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt
     7801 atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa
     7861 taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca
     7921 agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag
     7981 gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag
     8041 agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat
     8101 cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt
     8161 atatatttat attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac
     8221 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     8281 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     8341 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     8401 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     8461 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     8521 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     8581 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     8641 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgg
//
