LOCUS       sequence-263780-        9041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nucleo-cytoplasmic GFP in P-cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9041)
  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 9041)
  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: pAV0517
FEATURES             Location/Qualifiers
     source          1..9041
                     /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"
     primer_bind     2927..2943
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2928..2944
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5051..5764
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5081..5109)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5704..5724
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6781..6801)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6781..6799)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6820..6836)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6820..6836)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6833..6855)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6844..6860
                     /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(6868..6874,6875..6892,6893..6898))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6913..6934
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7051..7068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7222..7810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7302..7321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7981..8772,8773..8841))
                     /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     8604..8623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8842..8946)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9014..9032
                     /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 ggtaccgggc cccccctcga ggtcgacggt
     2941 atcgataagc ttgatatcga attcctgcag aagcatgcac gctgctcact tacaagataa
     3001 ccttttgttg aaaaggaaga tttattaact atatatgatt gaagattttt tcttttcaaa
     3061 ggatatataa gttctgcaaa gtttcctaaa aaaatctttc ttttttgttt cttacacctt
     3121 ttgacattta tattggagcc taacgttttg ccgctttgat atcatttatc cgttgatttt
     3181 agaaaaaaaa gttaggaaga gttgttaccg attttttcct tctttttatg ccaatattca
     3241 cttcatcaga tttaatttgg ctactttttt tttcgtaacc tactatagtt atctcattat
     3301 agaatgaaat gaaactctat tattatgtag ttaaaatcaa tgataagcac tgtattaaag
     3361 tttaagagaa aatcgcgaat tccttttgat gtccttttct caaaggatac aaacacgatt
     3421 tataacacct ttgcttcaga cgtttgattg atatgcctat tgtgatatac gaaacatcag
     3481 catcaggtta tagccgtgca acattttctg ataaatgatg aggaaaacag atgctattgt
     3541 cttgctcatg ttggttctct gggttgagat aatgaacata ttggtatata aaacttgcac
     3601 acggatcaac gagaattgct tacttttatt gctcttacat tttattgctt ttattttata
     3661 taacattgat tgctctcttc ttttcttgta ctaaaagaaa tacaaacata caaatatggc
     3721 ttacatgttt tacttcgcag aattataatt aattaaatac ccaagttatt cgttagcatg
     3781 gttgagtata ttgtttaata atagcgctat ttaagtcgta actttatagg ataagtatta
     3841 ccagtactaa tacgtgtaag aagttaccta gtgctattgt taacaaaata taagctcgtt
     3901 aatgtgagta acttgtttta ttttctactt gaaattttta acaaacaaac cagttaaaaa
     3961 aaattgatct ggatcaatac tccagctatt tggaaagaat tgacttacgc gtaaatgtgt
     4021 acgcgttaat gtttactgat ttcgttttcg ggtttagaga agtaaaggat ggaattagat
     4081 tacaggaaag agtattttta taaattatat ataaaggaag atatgactaa cggctagtgt
     4141 accaattcca taatataccg agtgcaattc aaaccaaact ctcgtatata ggctgcgatt
     4201 tataaagtaa aacttcagcg catcagcaca attgttcacg agtacaaaat ctaaaaagaa
     4261 attttctttg caatgcgctc atcaattaat ataaacgacc gcgatatatg aagtcaaaca
     4321 gtcgttaaac accaaaattt ctttacgtat ttacttcgca gattacccat caaaaagata
     4381 ttctatggat ttattaaaga taaagaattc gcttttgatg cctcaaattg gaatactatg
     4441 ctttagataa tcaaatgggc tgaactcttt gaccgaccga aaatggatct ctatgctgag
     4501 gcttctcacg gtaggtatat aaagagccta ctgcccttct tcaatatgcc aaataaatac
     4561 aaacaaggtt ctatactcaa cagtaatttt tcttactcat ttaattacga accttgctct
     4621 gttttcgcaa gttgttttca agttttcaag cttcgcgcgg gatatctggc gagcgatcaa
     4681 aggtgtttcc aatagcttta accttttcgt ttcctgtcct gtatttttaa ataatacatt
     4741 ttccgtgtct attactttta ttttggaaca atattgaagt tcaaattgta cacgactttg
     4801 gcaatcttag gaaaatcttg gcatatacaa ttccaatttt tgctctatcc aaatatcaag
     4861 actaccatca tcactgaatt tatactcttt tgtttgcgca tcgtctttcg gctttctttt
     4921 tcaccttctt taatattttg aaattctata taggtctttg tgtaccattt aatttttgta
     4981 tcttactttc ttttatatca tatttattag ttacttatac gttgagttta ccgtcgactc
     5041 tagaggatcc atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     5101 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
     5161 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
     5221 gccaacactt gtcactactt tcacttatgg tgttcaatgc ttttcaagat acccagatca
     5281 tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     5341 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
     5401 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     5461 tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca
     5521 aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca
     5581 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     5641 caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca
     5701 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     5761 caaacccggg taaaaggaat gtctcccttg ccagtactgc tagggttttt ctttcaaact
     5821 atggaagccc attcaagctg catattacga ttttgttttt cgcttttaga aagtggttta
     5881 gatgagataa tagaaaaatt cttgatctcc gacaacgagt acttttattt tttttgctaa
     5941 tcactttact caatattagc tcgaaatcgt agaaacgtag acgggtgcgg gataccgagt
     6001 ggtgtagtta agaattttta taaaccacgt ggcccaaaaa tatgaaccca aaacgtttat
     6061 acatgagtat actttaagaa ggctataccc cttcgtgtta gatgtagttt tagctaccca
     6121 acccgagtct atgagcttga cttcagatgt agaaggcatt aaatcgtttt gaatattaat
     6181 taaaaaacga tgaaaattaa atatttaaaa gcaatcatac gctgaaaatt tagtgctgtg
     6241 gctaatcctt caacatggaa atgccataaa agtgactttg acaaaaaaaa aagtatatac
     6301 aggtagtaaa ctcatctact tcattgactt tgtttacagc atgtggaagg aggaatattt
     6361 attgctaaat cgtagtttaa cattcaataa gtaatactat tgaaattcga caagattggc
     6421 cgcatggatg aaaaagaggc attttgcttt gggagaatta gtccaaatta gaactgaaaa
     6481 aaaaaacttt acgaggcaaa aatgtcggat tgagatcgta aaagttcgct cgtcgtcttt
     6541 tgctttgtga ttgttttcat ggatacatct tgctggatat ttaaatttta gtactatgta
     6601 taagatattc tataaatgtt ttatcaccca aacctgttag cgccttctta attctattca
     6661 atctggcttt tgctctgaga ctacttcttg gactttcact acttgttagt tatacggaat
     6721 ttgtgtaatt agaagtgaaa taatcctttc tattagtaat gcgagctcca gcttttgttc
     6781 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6841 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6901 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6961 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7021 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7081 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7141 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7201 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7261 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7321 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7381 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7441 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7501 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7561 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7621 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7681 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7741 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7801 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7921 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7981 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8041 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8101 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8161 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8221 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8281 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8341 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8401 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8461 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8521 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8581 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8641 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8701 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8761 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8821 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8881 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8941 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     9001 attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
