LOCUS       sequence-368000-        7163 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Swi6L-GFP with nuclear export signal mutated to alanines.
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 Apr 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: KBB416
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    70..91
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(106..111,112..129,130..136)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    144..160
                     /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     149..171
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     168..184
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     168..184
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        205..223
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     251..267
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     252..268
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1602..2312
                     /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(1629..1657)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2252..2272
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2363..2381)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2377..2424
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2631..2650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2632..2650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2657..2674)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2657..2673)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2666..2688)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2814..3269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2901..2920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3111..3132
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3400..4203)
                     /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(4204..4419)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4443..4462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4562..4584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4622..4640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4681..5184
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5221..5325
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5326..5394,5395..6186)
                     /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(5544..5563)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6357..6945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6846..6865
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7099..7116
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
       61 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
      121 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
      181 tgaccatgat tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggtacc
      241 gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcaggtcact
      301 gtcttctgct tcactctcgc cgccaatctc cttcttaaac gcttcaatag tatcaagaag
      361 ttctgtcgct agtgcggtca tctcttgcga ccgatcttct tcactcagta tcaattccac
      421 cgcatataac tgaagatagt atccgatgat ccctaaccca actttgtcaa aatccttcgc
      481 tgtagcgaca acccttgcag cgttactagc catagaattt atttaaaatg tggacaatta
      541 gtttctaaaa attgaatcaa ataaaataag aaagcttatt gttccttttt ttgtcttcat
      601 ctggaccgtt tggataagat cattcaccaa atcttagttt ttttttttag tttccttctc
      661 caacgatttc ataataataa aaaaaaacaa caaaaaacac gattgagaaa catcaatatg
      721 aactaagcaa tagctaaaaa aaaaagaata ataaagggga acacagtata attctcgaga
      781 ggatggcgtt ggaagaagtg gtacgatact taggacctca caatgagatc cctttgacac
      841 tcactagaga ctcggagact ggccatttcc tcctgaaaca ttttctgccc attttgcagc
      901 aatatcatga cacggggaat attaacgaga ccaaccccga tagtttcccc actgatgagg
      961 aaagaaataa attactggca cattatggaa ttgctgtaaa tacagacgac cgaggtgagt
     1021 tatggataga gctggaaaaa tgtttacaat tattaaacat gctaaattta ttcggtttgt
     1081 tccaggatgc attcgaattt gaggagcctg aaacagatca ggatgaagaa gatcccagcc
     1141 attcgaaact accagaaaac aagaccaaaa gtgaaaattc taaggataat atcagctcaa
     1201 agaggattaa taatttacaa gatatgagcc tggattctga tgcacacaga gaattaggct
     1261 ctcctttaaa aaaactaaaa atagatacct ctgtaataga tgctgagagt gactccactc
     1321 cgaatactgc cagaggcaag cctaacgatg atattaataa gggccctagc ggcgacaatg
     1381 aaaataatgg cactgatgac aatgacagaa ccgctggacc tatcataaca ttcactcatg
     1441 acctaacttc tgacttttta agcagtccac tgaaaatcat gaaagcacta ccttctccag
     1501 ttgtaaatga taatgaacag aagatgaaag cagaggcatt cgcacaacgg ttgttatttc
     1561 cagaaattca agaaatgcct acatccctta ataatgacag cagcaaagga gaagaactct
     1621 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     1681 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     1741 gcactactgg aaaactacct gttccatggc caacacttgt cactactttc acttatggtg
     1801 ttcagtgctt ttcaagatac ccggatcata tgaaacggca tgactttttc aagagtgcca
     1861 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     1921 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     1981 ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac tataactcac
     2041 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta
     2101 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     2161 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     2221 cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg
     2281 ggattacaca tggcatggat gaactataca aaggatccgg ctgctaacaa agcccgaaag
     2341 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2401 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac
     2461 gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac
     2521 tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat
     2581 caacgcatat agcgctagag cggccgccac cgcggtggag ctccaattcg ccctatagtg
     2641 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2701 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2761 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     2821 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2881 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2941 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3001 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3061 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3121 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3181 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3241 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     3301 tgcggtattt cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg
     3361 tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct
     3421 tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct
     3481 tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc
     3541 atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg
     3601 tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa
     3661 gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc
     3721 agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt
     3781 tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg
     3841 tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt
     3901 gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc
     3961 ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac
     4021 atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt
     4081 ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa
     4141 tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga
     4201 catgatttat cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg
     4261 gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc
     4321 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg
     4381 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa aggtggtatg gtgcactctc
     4441 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4501 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4561 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4621 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4681 gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa
     4741 tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga
     4801 aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca
     4861 aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat agatatacat
     4921 tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag
     4981 acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt
     5041 atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct
     5101 ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt
     5161 ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg
     5221 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5281 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5341 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5401 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5461 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5521 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5581 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5641 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5701 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5761 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5821 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5881 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5941 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6001 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6061 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6121 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6181 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6241 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6301 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6361 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6421 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6481 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6541 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6601 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6661 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6721 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6781 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6841 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6901 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6961 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7021 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7081 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7141 aaaccgcctc tccccgcgcg ttg
//
