LOCUS       sequence-368002-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Swi6L-GFP expressin N-term 273 amino acids of Swi6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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: KBB354
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(535..540,541..558,559..565)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    573..589
                     /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     578..600
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     597..613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     597..613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        634..652
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     680..696
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     681..697
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2031..2741
                     /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(2058..2086)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2681..2701
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2792..2810)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2806..2853
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3060..3079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3061..3079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3086..3103)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3086..3102)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3095..3117)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3243..3698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3330..3349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3540..3561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3829..4632)
                     /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(4633..4848)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4872..4891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4991..5013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5051..5069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5110..5613
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5650..5754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5755..5823,5824..6615)
                     /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(5973..5992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6786..7163,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      541 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      601 aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa gggaacaaaa
      661 gctggtaccg ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg
      721 caggtcactg tcttctgctt cactctcgcc gccaatctcc ttcttaaacg cttcaatagt
      781 atcaagaagt tctgtcgcta gtgcggtcat ctcttgcgac cgatcttctt cactcagtat
      841 caattccacc gcatataact gaagatagta tccgatgatc cctaacccaa ctttgtcaaa
      901 atccttcgct gtagcgacaa cccttgcagc gttactagcc atagaattta tttaaaatgt
      961 ggacaattag tttctaaaaa ttgaatcaaa taaaataaga aagcttattg ttcctttttt
     1021 tgtcttcatc tggaccgttt ggataagatc attcaccaaa tcttagtttt tttttttagt
     1081 ttccttctcc aacgatttca taataataaa aaaaaacaac aaaaaacacg attgagaaac
     1141 atcaatatga actaagcaat agctaaaaaa aaaagaataa taaaggggaa cacagtataa
     1201 ttctcgagag gatggcgttg gaagaagtgg tacgatactt aggacctcac aatgagatcc
     1261 ctttgacact cactagagac tcggagactg gccatttcct cctgaaacat tttctgccca
     1321 ttttgcagca atatcatgac acggggaata ttaacgagac caaccccgat agtttcccca
     1381 ctgatgagga aagaaataaa ttactggcac attatggaat tgctgtaaat acagacgacc
     1441 gaggtgagtt atggatagag ctggaaaaat gtttacaatt attaaacatg ctaaatttat
     1501 tcggtttgtt ccaggatgca ttcgaatttg aggagcctga aacagatcag gatgaagaag
     1561 atcccagcca ttcgaaacta ccagaaaaca agaccaaaag tgaaaattct aaggataata
     1621 tcagctcaaa gaggattaat aatttacaag atatgagcct ggattctgat gcacacagag
     1681 aattaggctc tcctttaaaa aaactaaaaa tagatacctc tgtaatagat gctgagagtg
     1741 actccactcc gaatactgcc agaggcaagc ctaacgatga tattaataag ggccctagcg
     1801 gcgacaatga aaataatggc actgatgaca atgacagaac cgctggacct atcataacat
     1861 tcactcatga cctaacttct gactttttaa gcagtccact gaaaatcatg aaagcactac
     1921 cttctccagt tgtaaatgat aatgaacaga agatgaaact agaggcattc ttacaacggt
     1981 tgttatttcc agaaattcaa gaaatgccta catcccttaa taatgacagc agcaaaggag
     2041 aagaactctt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     2101 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     2161 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttca
     2221 cttatggtgt tcagtgcttt tcaagatacc cggatcatat gaaacggcat gactttttca
     2281 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
     2341 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     2401 taaaaggtat tgattttaaa gaagatggaa acattcttgg gcacaaattg gaatacaact
     2461 ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagctaact
     2521 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     2581 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat
     2641 ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     2701 cagctgctgg gattacacat ggcatggatg aactatacaa aggatccggc tgctaacaaa
     2761 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2821 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat
     2881 ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc
     2941 gagcaggact gggcggcggc caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag
     3001 aaattgcatc aacgcatata gcgctagagc ggccgccacc gcggtggagc tccaattcgc
     3061 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3121 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3181 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3241 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3301 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3361 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3421 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3481 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3541 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3601 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3661 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     3721 acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc
     3781 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     3841 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     3901 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     3961 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     4021 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     4081 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     4141 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     4201 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     4261 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     4321 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     4381 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     4441 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     4501 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     4561 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     4621 agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa
     4681 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     4741 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     4801 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg
     4861 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4921 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4981 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5041 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5101 tcttagacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
     5161 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
     5221 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
     5281 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
     5341 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
     5401 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
     5461 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
     5521 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
     5581 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
     5641 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5701 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5761 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5821 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5881 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5941 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6001 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6061 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6121 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6181 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6241 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6301 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6361 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6421 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6481 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6541 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6601 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6661 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6721 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6781 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6841 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6901 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6961 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7021 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7081 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7141 acgacctaca ccgaactgag ata
//
