LOCUS       sequence-368085-        6305 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Smaller fusion of Nma111 nuclear localization signal to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6305)
  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 6305)
  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: KBB282
FEATURES             Location/Qualifiers
     source          1..6305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        141..245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(246..314,315..1106)
                     /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(464..483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1277..1865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1766..1785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2019..2036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2153..2174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2189..2194,2195..2212,2213..2219)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2227..2243
                     /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     2232..2254
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2251..2267
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2251..2267
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2288..2306
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2334..2350
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2335..2351
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2827..3537
                     /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(2854..2882)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3477..3497
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3588..3606)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3602..3649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3856..3875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3857..3875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3882..3899)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3882..3898)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3891..3913)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4039..4494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4126..4145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4336..4357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4625..5428)
                     /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(5429..5644)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5668..5687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5787..5809
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5847..5865)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5906..6305,1..104)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt
       61 tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg
      121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      481 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      781 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      901 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2041 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2101 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2161 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2221 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2281 agctcgaaat taaccctcac taaagggaac aaaagctggt accgggcccc ccctcgaggt
     2341 cgacggtatc gataagcttg atatcgaatt cctgcaggca tcagcatcag caagatccaa
     2401 gaacctagag tcacgattga aatatttaat aataaatgct atgtatcata gataaactag
     2461 aaagaggcaa gaaaaattta acaagtttct taattgaata ttattttact gtcactacca
     2521 ttattagacg ccatctcagc gtgtgacatg ataatttttc tttttttctt ttttttcctc
     2581 aaggcgatga tatgaaactt tcaaaaaatt tatatgtaaa taaaaaagtt ctacgagggt
     2641 ttaaatacgg aagtttattt agaagatatt tatactctga atacacacgt agagtacagt
     2701 aaaggttttt tagatctact aatgaccata tcgttgagca atataaagaa aagagaccat
     2761 tctaaaattt ccgatggaac ttctggtgaa tcatcgcttg taaaaaggaa acagttggaa
     2821 agcgccagca aaggagaaga actcttcact ggagttgtcc caattcttgt tgaattagat
     2881 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
     2941 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca
     3001 cttgtcacta ctttcactta tggtgttcag tgcttttcaa gatacccgga tcatatgaaa
     3061 cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
     3121 ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt
     3181 gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttgggcac
     3241 aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat
     3301 ggaatcaaag ctaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca
     3361 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
     3421 tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc
     3481 cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaagga
     3541 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     3601 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     3661 aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg
     3721 ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga
     3781 gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagagcggcc gccaccgcgg
     3841 tggagctcca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     3901 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     3961 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4021 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4081 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4141 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4201 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4261 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4321 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4381 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4441 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     4501 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat
     4561 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     4621 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
     4681 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
     4741 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
     4801 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
     4861 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
     4921 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
     4981 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
     5041 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
     5101 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
     5161 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
     5221 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
     5281 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
     5341 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
     5401 cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc
     5461 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     5521 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     5581 aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat
     5641 tgaaaaggtg gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5701 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5761 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5821 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5881 catgataata atggtttctt agacggatcg cttgcctgta acttacacgc gcctcgtatc
     5941 ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat
     6001 ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata
     6061 aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag
     6121 aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga
     6181 ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag
     6241 caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct
     6301 tcgga
//
