LOCUS       sequence-263743-        8266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sfGFP excluded from the nucleus with two NES signals .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8266)
  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 8266)
  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: pAV0479
FEATURES             Location/Qualifiers
     source          1..8266
                     /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"
     gene            4830..5949
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        4830..5173
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5174..5746
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      5752..5949
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6006..6026)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6006..6024)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6045..6061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6045..6061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6058..6080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6069..6085
                     /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(6093..6099,6100..6117,6118..6123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6138..6159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6276..6293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6447..7035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6527..6546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7206..7997,7998..8066))
                     /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     7829..7848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8067..8171)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8239..8257
                     /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 ttcccaaccc ttgtcttgct
     3961 cattgagaca gctgtctatt tctccttcca agggtgaaga gctatttact ggggttgtac
     4021 ccattttggt agaactggac ggagatgtaa acggacataa attctctgtt agaggtgagg
     4081 gcgaaggcga tgccaccaat ggtaaattga ctctgaagtt tatatgcact acgggtaaat
     4141 tacctgttcc ttggccaacc ctagtaacaa ctttgacata tggtgttcaa tgtttctcaa
     4201 gatacccaga ccatatgaaa aggcatgatt tctttaaaag tgctatgcca gaaggctacg
     4261 tgcaagagag aactatctcc tttaaggatg acggtacgta taaaacacga gcagaagtga
     4321 aattcgaagg ggatacacta gttaatcgca tcgaattaaa gggtatagac tttaaggaag
     4381 atggtaatat tctcggccat aaacttgagt ataatttcaa ctcgcataat gtgtacatta
     4441 cagctgacaa acaaaagaac ggaattaaag cgaattttaa aatcaggcac aacgtcgaag
     4501 atgggtctgt tcaacttgcc gatcattatc agcaaaacac ccctattggt gatggtccag
     4561 tcttgttacc cgataatcac tacttaagca cacagtctag attgtcaaaa gatccgaatg
     4621 aaaagcgtga tcacatggtt ttattggaat ttgtcaccgc tgcaggaata actcacggaa
     4681 tggacgagct ttataaggag gacttggtaa ttgcaatgga tcaactgaat ttggagcaag
     4741 gtggtcccgg tggtcatgga tccccgggtt aattaaggcg cgccagatct gtttagcttg
     4801 cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca
     4861 gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac
     4921 atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa
     4981 attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga
     5041 attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt
     5101 tcttctttca tatacttcct ttttaaatct tgctaggata cagttctcac atcacatccg
     5161 aacataaaca accatgggta ccactcttga cgacacggct taccggtacc gcaccagtgt
     5221 cccgggggac gccgaggcca tcgaggcact ggatgggtcc ttcaccaccg acaccgtctt
     5281 ccgcgtcacc gccaccgggg acggcttcac cctgcgggag gtgccggtgg acccgcccct
     5341 gaccaaggtg ttccccgacg acgaatcgga cgacgaatcg gacgacgggg aggacggcga
     5401 cccggactct cggacgttcg tcgcgtacgg ggacgacggc gacctggcgg gcttcgtggt
     5461 cgtctcgtac tccggctgga accgccggct gaccgtcgag gacatcgagg tcgccccgga
     5521 gcaccggggg cacggggtcg ggcgcgcgtt gatggggctc gcgacggagt tcgcccgcga
     5581 gcggggtgcc gggcacctct ggctggaggt caccaacgtc aacgcaccgg cgatccacgc
     5641 gtaccggcgg atggggttca ccctctgcgg cctggacacc gccctgtacg acggcaccgc
     5701 ctcggacggc gagcaggcgc tctacatgag catgccctgc ccctaatcag tactgacaat
     5761 aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt
     5821 ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca
     5881 gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc
     5941 gctatactgc tgtcgattcg atactaacgc cgccatccag tttaaacgag ctccagcttt
     6001 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     6061 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6121 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6181 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6241 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6301 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6361 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6421 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6481 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6541 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6601 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6661 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6721 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6781 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6841 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6901 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6961 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7021 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7081 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7141 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7201 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7261 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7321 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7381 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7441 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7501 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7561 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7621 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7681 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7741 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7801 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7861 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7921 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7981 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8041 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8101 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8161 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8221 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
