LOCUS       sequence-368133-        6450 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Larger fusion of Nma111 nuclear localization signal to GFP .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  AUTHORS   Belanger KD, Walter D, Henderson TA, Yelton AL, O'Brien TG, Belanger
            KG, Geier SJ, Fahrenkrog B
  TITLE     Nuclear localisation is crucial for the proapoptotic activity of the
            HtrA-like serine protease Nma111p.
  JOURNAL   J Cell Sci. 2009 Nov 1;122(Pt 21):3931-41. doi: 10.1242/jcs.056887. 
            Epub 2009 Oct 6.
  PUBMED    19808885
REFERENCE   2  (bases 1 to 6450)
  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: "J Cell 
            Sci."; date: "2009-11-1"; volume: "122(Pt 21):3931-41. doi"; pages: 
            " 10.1242/jcs.056887. Epub 2009 Oct 6"
COMMENT     Sequence Label: KBB280
FEATURES             Location/Qualifiers
     source          1..6450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        380..484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(485..553,554..1345)
                     /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(703..722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1516..2104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2005..2024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2258..2275
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2392..2413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2428..2433,2434..2451,2452..2458)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2466..2482
                     /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     2471..2493
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2490..2506
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2490..2506
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2527..2545
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2573..2589
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2574..2590
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3211..3921
                     /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(3238..3266)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3861..3881
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3972..3990)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3986..4033
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4240..4259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4241..4259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4266..4283)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4266..4282)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4275..4297)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4423..4878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4510..4529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4720..4741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5009..5812)
                     /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(5813..6028)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6052..6071)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6171..6193
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6231..6249)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(6290..6450,1..343)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc
       61 agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg
      121 tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa
      181 gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa
      241 aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat ttatatattt
      301 atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg
      361 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      421 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      481 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      541 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      601 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      661 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      721 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      781 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      841 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      901 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      961 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1021 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1081 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1141 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1201 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1261 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1321 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1381 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1441 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1621 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2221 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2281 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2341 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     2401 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     2461 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     2521 gctcgaaatt aaccctcact aaagggaaca aaagctggta ccgggccccc cctcgaggtc
     2581 gacggtatcg ataagcttga tatcgaattc ctgcaggcat cagcatcagc aagatccaag
     2641 aacctagagt cacgattgaa atatttaata ataaatgcta tgtatcatag ataaactaga
     2701 aagaggcaag aaaaatttaa caagtttctt aattgaatat tattttactg tcgctaccat
     2761 tattagacgc catctcagcg tgtgacatga taatttttct ttttttcttt ttttttcctc
     2821 aaggcgatga tatgaaactt tcaaaaaatt tatatgtaaa taaaaaagtt ctacgagggt
     2881 ttaaatacgg aagtttattt agaagatatt tatactctga atacacacgt agagtacagt
     2941 aaaggttttt tagatctact aatgaccata tcgttgagca atataaagaa aagagaccat
     3001 tctaaaattt ccgatggaac ttctggtgaa tcatcgcttg taaaaaggaa acagttggaa
     3061 agcgccaccg gagatcaaga ggaagaatat acagaccatg aaatcattat agagccgttg
     3121 cactttgcta ataataataa cactgtactc acagattcag aaaattatct aagatggcaa
     3181 aataccatct ccaacgttgt taaatcagtg agcaaaggag aagaactctt cactggagtt
     3241 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
     3301 gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga
     3361 aaactacctg ttccatggcc aacacttgtc actactttca cttatggtgt tcagtgcttt
     3421 tcaagatacc cggatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt
     3481 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa
     3541 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
     3601 gaagatggaa acattcttgg gcacaaattg gaatacaact ataactcaca caatgtatac
     3661 atcatggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattag acacaacatt
     3721 gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     3781 cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc
     3841 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat
     3901 ggcatggatg aactatacaa aggatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3961 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     4021 gaggggtttt ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg
     4081 ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc
     4141 caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata
     4201 gcgctagagc ggccgccacc gcggtggagc tccaattcgc cctatagtga gtcgtattac
     4261 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4321 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4381 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     4441 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4501 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4561 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4621 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4681 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4741 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4801 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4861 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4921 acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     4981 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     5041 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     5101 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     5161 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     5221 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     5281 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
     5341 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     5401 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     5461 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     5521 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     5581 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
     5641 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
     5701 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
     5761 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
     5821 ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
     5881 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     5941 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
     6001 aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg tgcactctca gtacaatctg
     6061 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     6121 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     6181 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     6241 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgg atcgcttgcc
     6301 tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg
     6361 tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag
     6421 ttacggaatg aagaaaaaaa aataaacaaa
//
