LOCUS       sequence-367443-        6889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Swi6M-GFP expressing N-term 181 amino acids of Swi6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6889)
  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 6889)
  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: KBB353
FEATURES             Location/Qualifiers
     source          1..6889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        426..530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(531..599,600..1391)
                     /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(749..768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1562..2150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2051..2070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2304..2321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2438..2459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2474..2479,2480..2497,2498..2504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2512..2528
                     /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     2517..2539
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2536..2552
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2536..2552
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2573..2591
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2619..2635
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2620..2636
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3696..4406
                     /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="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3723..3751)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4346..4366
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4457..4475)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4471..4518
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4725..4744)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4726..4744)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4751..4768)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4751..4767)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4760..4782)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4908..5363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4995..5014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5205..5226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5494..6297)
                     /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(6298..6513)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6537..6556)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6656..6678
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6716..6734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(6775..6889,1..389)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt
       61 caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat taaatagata
      121 tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta
      181 cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag
      241 gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt
      301 tttctttaat ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt
      361 aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact tttcggggaa
      421 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      481 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      541 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      601 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
      661 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
      721 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
      781 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
      841 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
      901 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
      961 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1021 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1081 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1141 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1201 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1261 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1321 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1381 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1441 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1501 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1561 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1681 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1921 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2161 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2221 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2281 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2341 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2401 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     2461 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     2521 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc
     2581 ctcactaaag ggaacaaaag ctggtaccgg gccccccctc gaggtcgacg gtatcgataa
     2641 gcttgatatc gaattcctgc aggtcactgt cttctgcttc actctcgccg ccaatctcct
     2701 tcttaaacgc ttcaatagta tcaagaagtt ctgtcgctag tgcggtcatc tcttgcgacc
     2761 gatcttcttc actcagtatc aattccaccg catataactg aagatagtat ccgatgatcc
     2821 ctaacccaac tttgtcaaaa tccttcgctg tagcgacaac ccttgcagcg ttactagcca
     2881 tagaatttat ttaaaatgtg gacaattagt ttctaaaaat tgaatcaaat aaaataagaa
     2941 agcttattgt tccttttttt gtcttcatct ggaccgtttg gataagatca ttcaccaaat
     3001 cttagttttt ttttttttag tttccttctc caacgatttc ataataataa aaaaaaacaa
     3061 caaaaaacac gattgagaaa catcaatatg aactaagcaa tagctaaaaa aaaaagaata
     3121 ataaagggga acacagtata attctcgaga ggatggcgtt ggaagaagtg gtacgatact
     3181 taggacctca caatgagatc cctttgacac tcactagaga ctcggagact ggccatttcc
     3241 tcctgaaaca ttttctgccc attttgcagc aatatcatga cacggggaat attaacgaga
     3301 ccaaccccga tagtttcccc actgatgagg aaagaaataa attactggca cattatggaa
     3361 ttgctgtaaa tacagacgac cgaggtgagt tatggataga gctggaaaaa tgtttacaat
     3421 tattaaacat gctaaattta ttcggtttgt tccaggatgc attcgaattt gaggagcctg
     3481 aaacagatca ggatgaagaa gatcccagcc attcgaaact accagaaaac aagaccaaaa
     3541 gtgaaaattc taaggataat atcagctcaa agaggattaa taatttacaa gatatgagcc
     3601 tggattctga tgcacacaga gaattaggct ctcctttaaa aaaactaaaa atagatacct
     3661 ctgtaataga tgctgagagt gactccactc cgaatagcaa aggagaagaa ctcttcactg
     3721 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     3781 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     3841 ctggaaaact acctgttcca tggccaacac ttgtcactac tttcacttat ggtgttcagt
     3901 gcttttcaag atacccggat catatgaaac ggcatgactt tttcaagagt gccatgcccg
     3961 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     4021 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     4081 ttaaagaaga tggaaacatt cttgggcaca aattggaata caactataac tcacacaatg
     4141 tatacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     4201 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     4261 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     4321 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     4381 cacatggcat ggatgaacta tacaaaggat ccggctgcta acaaagcccg aaaggaagct
     4441 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     4501 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca ggacgggtgt
     4561 ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca ggactgggcg
     4621 gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt gcatcaacgc
     4681 atatagcgct agagcggccg ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt
     4741 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4801 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4861 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     4921 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4981 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5041 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5101 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5161 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5221 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5281 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5341 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     5401 atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt
     5461 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
     5521 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
     5581 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     5641 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     5701 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     5761 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     5821 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     5881 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     5941 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     6001 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     6061 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     6121 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     6181 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     6241 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     6301 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
     6361 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     6421 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca
     6481 aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca
     6541 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     6601 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     6661 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     6721 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacggatcgc
     6781 ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact
     6841 ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaat
//
