LOCUS       sequence-263631-        8227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sfGFP targeted to the nucleus with two NLS signals .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8227)
  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 8227)
  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: pAV0478
FEATURES             Location/Qualifiers
     source          1..8227
                     /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             3945..3965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4680..4700
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     gene            4791..5910
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        4791..5134
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5135..5707
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      5713..5910
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5967..5987)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5967..5985)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6006..6022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6006..6022)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6019..6041)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6030..6046
                     /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(6054..6060,6061..6078,6079..6084))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6099..6120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6237..6254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6408..6996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6488..6507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7167..7958,7959..8027))
                     /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     7790..7809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8028..8132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8200..8218
                     /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 ttcccctaag aagaagcgta
     3961 aggtttccaa gggtgaagag ctatttactg gggttgtacc cattttggta gaactggacg
     4021 gagatgtaaa cggacataaa ttctctgtta gaggtgaggg cgaaggcgat gccaccaatg
     4081 gtaaattgac tctgaagttt atatgcacta cgggtaaatt acctgttcct tggccaaccc
     4141 tagtaacaac tttgacatat ggtgttcaat gtttctcaag atacccagac catatgaaaa
     4201 ggcatgattt ctttaaaagt gctatgccag aaggctacgt gcaagagaga actatctcct
     4261 ttaaggatga cggtacgtat aaaacacgag cagaagtgaa attcgaaggg gatacactag
     4321 ttaatcgcat cgaattaaag ggtatagact ttaaggaaga tggtaatatt ctcggccata
     4381 aacttgagta taatttcaac tcgcataatg tgtacattac agctgacaaa caaaagaacg
     4441 gaattaaagc gaattttaaa atcaggcaca acgtcgaaga tgggtctgtt caacttgccg
     4501 atcattatca gcaaaacacc cctattggtg atggtccagt cttgttaccc gataatcact
     4561 acttaagcac acagtctaga ttgtcaaaag atccgaatga aaagcgtgat cacatggttt
     4621 tattggaatt tgtcaccgct gcaggaataa ctcacggaat ggacgagctt tataaggggc
     4681 ctaagaagaa gcgtaaggtt ggtggtcccg gtggtcatgg atccccgggt taattaaggc
     4741 gcgccagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg
     4801 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
     4861 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
     4921 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg
     4981 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     5041 gttaggattt gccactgagg ttcttctttc atatacttcc tttttaaatc ttgctaggat
     5101 acagttctca catcacatcc gaacataaac aaccatgggt accactcttg acgacacggc
     5161 ttaccggtac cgcaccagtg tcccggggga cgccgaggcc atcgaggcac tggatgggtc
     5221 cttcaccacc gacaccgtct tccgcgtcac cgccaccggg gacggcttca ccctgcggga
     5281 ggtgccggtg gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc
     5341 ggacgacggg gaggacggcg acccggactc tcggacgttc gtcgcgtacg gggacgacgg
     5401 cgacctggcg ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga
     5461 ggacatcgag gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct
     5521 cgcgacggag ttcgcccgcg agcggggtgc cgggcacctc tggctggagg tcaccaacgt
     5581 caacgcaccg gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac
     5641 cgccctgtac gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg
     5701 cccctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     5761 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     5821 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     5881 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca
     5941 gtttaaacga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     6001 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6061 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6121 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6181 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6241 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6301 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6361 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6421 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6841 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8161 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8221 tttcgtc
//
