LOCUS       sequence-263685-        8203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sfGFP targeted to the nucleus with one NLS signals .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8203)
  AUTHORS   Vjestica A, Marek M, Nkosi PJ, Merlini L, Liu G, Berard M, 
            Billault-Chaumartin I, Martin SG
  TITLE     A toolbox of stable integration vectors in the fission yeast 
            Schizosaccharomyces pombe.
  JOURNAL   J Cell Sci. 2020 Jan 8;133(1). pii: jcs.240754. doi: 
            10.1242/jcs.240754.
  PUBMED    31801797
REFERENCE   2  (bases 1 to 8203)
  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: "J Cell 
            Sci."; date: "2020-01-8"; volume: "133(1). pii: jcs.240754. doi"; 
            pages: " 10.1242/jcs.240754"
COMMENT     Sequence Label: pAV0526
FEATURES             Location/Qualifiers
     source          1..8203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             1178..1828
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQK
                     GEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQA
                     MGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLYS
                     FSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     rep_origin      complement(2262..2717)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2399..2420)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2611..2630
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2843..2865
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2857..2874
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2858..2874
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2884..2903
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2884..2902
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4656..4676
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     gene            4767..5886
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        4767..5110
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5111..5683
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      5689..5886
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5943..5963)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5943..5961)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5982..5998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5982..5998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5995..6017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6006..6022
                     /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)."
     promoter        complement(join(6030..6036,6037..6054,6055..6060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6075..6096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6213..6230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6384..6972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6464..6483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7143..7934,7935..8003))
                     /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     7766..7785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8004..8108)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8176..8194
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaccg ggatggaacg attgaatact gtgctactct ttatgtaata taacgaattc
      241 aactgttcaa cgtatgtgac tgctctctgc ttctcggtat atatttactt gaaggaatta
      301 atttttcttt gccaatggtc tcttttacca attgcagtct ggcttaaagc aatgaggttg
      361 taattaaatg gcttgagacc tagaacaaac accttttaca atccttgttt actggttttg
      421 atagtaaatt cggttcgctt gtggtgtaca ttatattttt taggctagtc gtgatttttt
      481 gataatatca atattcttag atttgagaca ttcatgcttt ttaatttgat caatcgtttg
      541 tatcttcatt aattagtagg tggtttgata gaacatttag ttgcttgttt gactacaaat
      601 atctgcagaa tggattcggt taataaaaaa aatgtttgag gaattaattt tcgctttgga
      661 gtggaggcct caattcgatg ttgcagattt tacaagtttt taaaatgtat ttcattatta
      721 ctttttatat gcctaataaa aaagccatag tttaatctat agataacttt ttttccagtg
      781 cactaacgga cgttacattc ccatacaaaa ctgcgtagtt aaagctaagg aaaagttaat
      841 atcatgttaa ttaaatacgc tatttacaat aagacattga actcatttat atcgttgaat
      901 atgaataacc aatttcagcg aatttttaac aaacatcgtt cacctcgttt aaggatatct
      961 tgtgtatggg gtgttgactt gctttatcga ataattaccg tacctgtaat tggcttgctg
     1021 gatatagcgg tagtctaata tctagcaaaa atcttttggg tgaaaaggct tgcaatttca
     1081 cgacaccgaa ctatttgtca ttttttaata aggaagtttt ccataaattc ctgtaattct
     1141 cggttgatct aattgaaaag agtagttttg catcacgatg aggagggctt ttgtagaaag
     1201 aaatacgaac gaaacgaaaa tcagcgttgc catcgctttg gacaaagctc ccttacctga
     1261 agagtcgaat tttattgatg aacttataac ttccaagcat gcaaaccaaa agggagaaca
     1321 agtaatccaa gtagacacgg gaattggatt cttggatcac atgtatcatg cactggctaa
     1381 acatgcaggc tggagcttac gactttactc aagaggtgat ttaatcatcg atgatcatca
     1441 cactgcagaa gatactgcta ttgcacttgg tattgcattc aagcaggcta tgggtaactt
     1501 tgccggcgtt aaaagatttg gacatgctta ttgtccactt gacgaagctc tttctagaag
     1561 cgtagttgac ttgtcgggac ggccctatgc tgttatcgat ttgggattaa agcgtgaaaa
     1621 ggttggggaa ttgtcctgtg aaatgatccc tcacttacta tattcctttt cggtagcagc
     1681 tggaattact ttgcatgtta cctgcttata tggtagtaat gaccatcatc gtgctgaaag
     1741 cgcttttaaa tctctggctg ttgccatgcg cgcggctact agtcttactg gaagttctga
     1801 agtcccaagc acgaagggag tgttgtaaag atgaattgga ttatgtcagg aaaagaacga
     1861 caattttgca tccaaattgt ctaaatttta gagttgcttg aaaacaatag aaccttactt
     1921 gctttataat tacgttaatt agaagcgtta tctcgtgaag gaatatagta cgtagccgta
     1981 taaattgaat tgaatgttca gcttatagaa tagagacact ttgctgttca atgcgtcgtc
     2041 acttaccata ctcactttat tatacgactt taagtataaa ctccgcggtt atggtaaaat
     2101 taatgatgca caaacgtccg attccatatg ggtacactac aattaaatac ttttaagtaa
     2161 cgctgatttc ttatttttta tccttggtgt atttattgtt tataccgcta gggcggtgtg
     2221 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     2281 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2341 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2401 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2461 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2521 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2581 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2641 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2701 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2761 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     2821 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     2881 gcgtaatacg actcactata gggcgaattg ggtacctaaa gtatggaaaa tcaaaattcc
     2941 tgaatttttg taaactccca tacaaatacg attacacata catacaaaaa tgcaaataac
     3001 aaaatataca cgcacgcatg cacactaaat aaacaaactg tgcgtccatt ccttttatgc
     3061 acaaacgttg aagctcagga catcgcacca acaatcactc atctcttctc tctctaccca
     3121 ctttccaccc acacccacag tctgcatctg tttgacgact gtagctcaag catcagtagc
     3181 agccgctgtc atcctcaact gctgctgctg atgctgcttg aatgtacgca tgatagcaac
     3241 gacgcaccgc tctaccacta ctaccgaagc gccatcatca cctcctcacc taccatcgca
     3301 atcgtcctct cgaaactctt cgaccgcttt cacccggctt cctgcaggta acaatgcgct
     3361 cggcgcttga ttaaaagagg ccagaggcga taaatgactc agggtgcatt tggcctcgct
     3421 acgaaaggac cctgtcttgt gatttttccg tagccaatgg acaaccgcct tgtcaacctt
     3481 ttgactttgg aatgtgctag caattcctcc ttccccctga cttcaggtga caaggcttgc
     3541 atcacgctga ggtaggcgtc ggaaaaactt tcctcacaag gctgacgtca tggaatgtgc
     3601 aatcaaagtc aggagcggga acgaggcaga ttaccaacga agcaacgacc atctcggaac
     3661 aacttgcaaa gaaagtcaag gaaagttggc accacatcac gatcgcgcac cttctccaat
     3721 caccttgctc cttagttgtt ccgaatcgag tgcgaggcaa ttgtggtaaa gatagtcggg
     3781 ggaaaccctt accccattca caccacggct gatggctgct gtgtgggatg gaaaggtata
     3841 aatagaggca ggcggtgcct agcatcgctt ctgtatagat cattcatcca tagtattgat
     3901 ttacacttga ttcaaaccta ggccgcggaa gggggatggg ttccaagggt gaagagctat
     3961 ttactggggt tgtacccatt ttggtagaac tggacggaga tgtaaacgga cataaattct
     4021 ctgttagagg tgagggcgaa ggcgatgcca ccaatggtaa attgactctg aagtttatat
     4081 gcactacggg taaattacct gttccttggc caaccctagt aacaactttg acatatggtg
     4141 ttcaatgttt ctcaagatac ccagaccata tgaaaaggca tgatttcttt aaaagtgcta
     4201 tgccagaagg ctacgtgcaa gagagaacta tctcctttaa ggatgacggt acgtataaaa
     4261 cacgagcaga agtgaaattc gaaggggata cactagttaa tcgcatcgaa ttaaagggta
     4321 tagactttaa ggaagatggt aatattctcg gccataaact tgagtataat ttcaactcgc
     4381 ataatgtgta cattacagct gacaaacaaa agaacggaat taaagcgaat tttaaaatca
     4441 ggcacaacgt cgaagatggg tctgttcaac ttgccgatca ttatcagcaa aacaccccta
     4501 ttggtgatgg tccagtcttg ttacccgata atcactactt aagcacacag tctagattgt
     4561 caaaagatcc gaatgaaaag cgtgatcaca tggttttatt ggaatttgtc accgctgcag
     4621 gaataactca cggaatggac gagctttata aggggcctaa gaagaagcgt aaggttggtg
     4681 gtcccggtgg tcatggatcc ccgggttaat taaggcgcgc cagatctgtt tagcttgcct
     4741 cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc
     4801 ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc
     4861 cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt
     4921 acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt
     4981 gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct
     5041 tctttcatat acttcctttt taaatcttgc taggatacag ttctcacatc acatccgaac
     5101 ataaacaacc atgggtacca ctcttgacga cacggcttac cggtaccgca ccagtgtccc
     5161 gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca ccgtcttccg
     5221 cgtcaccgcc accggggacg gcttcaccct gcgggaggtg ccggtggacc cgcccctgac
     5281 caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg acggcgaccc
     5341 ggactctcgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct tcgtggtcgt
     5401 ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg ccccggagca
     5461 ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg cccgcgagcg
     5521 gggtgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga tccacgcgta
     5581 ccggcggatg gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg gcaccgcctc
     5641 ggacggcgag caggcgctct acatgagcat gccctgcccc taatcagtac tgacaataaa
     5701 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     5761 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     5821 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     5881 atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc cagcttttgt
     5941 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     6001 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6061 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6121 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6181 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6241 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6301 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6361 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6421 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6481 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6541 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6601 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6661 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6721 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6781 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6841 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6901 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6961 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7021 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7081 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7141 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7201 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7261 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7321 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7381 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7441 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7501 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7561 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7621 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7681 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7741 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7801 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7861 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7921 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7981 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8101 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8161 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
