LOCUS       sequence-198903-        8133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt41 promoter with Sp.ura4 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8133)
  AUTHORS   Zhao Y, Boeke JD
  TITLE     Construction of Designer Selectable Marker Deletions with 
            CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized 
            Design of Common Entry Vectors.
  JOURNAL   G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 
            10.1534/g3.117.300363.
  PUBMED    29321167
REFERENCE   2  (bases 1 to 8133)
  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: "G3 
            (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 
            10.1534/g3.117.300363."
COMMENT     Sequence Label: pYZ156
FEATURES             Location/Qualifiers
     source          1..8133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             217..597
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISRLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    273..380
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(388..407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(389..407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(417..434)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(417..433)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(426..448)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      574..1029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(661..680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     871..892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1465..2259)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     promoter        2818..3972
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      5069..5852
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6228..6332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6333..6401,6402..7193)
                     /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(6551..6570)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7364..7952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7853..7872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8106..8123
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta
      301 gaactagtcg atccgccggg ctgcaggaat tcgatatcaa ggttatcgat accgtccacc
      361 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
      421 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      481 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      541 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
      601 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      661 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      721 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      781 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      841 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      901 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      961 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     1021 ttacaattta gcttgtgata ttgacgaaac tttttgacat ctaatttatt ctgttccaac
     1081 accaatgttt ataaccaagt tttatcttgt ttgtctacat ggtattttac attcatctac
     1141 atacatcttt cattggcttt gtacatagtt atcattacaa gtctaaaaaa attcactctt
     1201 ttcttattca atgtcaatcc aagagaaaag attgtggtaa tgttgtagga gcatgtttaa
     1261 taaattacta tagcaaatta ctttttattc ccaaggtgtt tatctataat agttaatatt
     1321 ttagtcgcta cataaaattt taccaaagag tacttgtata ctaattctaa atgccttctg
     1381 acataaaacg cctaggaaaa caaacgcaaa caaggcatcg actttttcaa taaccaacca
     1441 aaaaaatttt acattagtct ttttttaatg ctgagaaagt ctttgctgat atgccttcca
     1501 accagcttct ctatatctct tggcttcgac aacaggatta cgaccagctc catagactcc
     1561 acgaccaaca atgatgatat cgctaccgca gtttacaatc acttcttcag gagtacgata
     1621 ttgctgtccc agcccgtctc ctttaacatc caagccgata ccaggggaca tagttatgta
     1681 gtcgctttga aggttaggaa atcgacgacc agctataaag ccaaagcaaa aatcggtatg
     1741 cttctcaaac cattctaagg ttttctctgt gtaggaacca gtagccaaag agcctttgga
     1801 agacatttca gccaaaagca agagtccacg tcccaaaggt aaaccaactt ctttgaggcc
     1861 ttgtataata ccctcgcctg gcactgtatg gcaatttgtg atatgagccc aagaagcaat
     1921 tttgtacaca ccagatgcat attgtagctt gacggtattt ccaatgtctg cgaatttgcg
     1981 atcctcaaag ataagaaaac gatgcttttt acctaaggcc accagttttt ctaccatatc
     2041 ctggtcgaaa tcctcgacaa cgtcaatatg tgtcttgata acacagacat agggtccaat
     2101 tttatctacc aattctaaga tttcggattt cttcgtcaaa tcgaccgcga ctgacaagtt
     2161 gctttgcttt tcttccatca aagccaacaa ttccttggca atgggatttt tcatcccctc
     2221 agctctagct gaatagcttt gaaatactct agcatccata actttgcttt taaaccttta
     2281 atttcgatcc aagcaaaaaa gaggttcttg gtaggacaat acggtaagaa aacacgacat
     2341 gtgcagagat gccgacgaag catagttaaa ctgggatggt aaaatcaatt aagaatttat
     2401 aaagacaaaa ttgtataagt ctctaaaaca tcttaattat acctcacaga actatctaaa
     2461 atatattcac aaagtgcaaa cattatcatg aaaaagaacc attttaattt aaagcaaggg
     2521 cattaaggct tatttacaga atttcttact tttgtaaaga ttataaggct gattatcttt
     2581 ttcaccatgc caaaaattac acaagataga atggatgttt gaaattaaac gtgagtatac
     2641 aaacaaatac actaggtaaa tcgaaacatt tttttctcca ttaagtaaca aattcctatt
     2701 tagagaaaga atgctgagta gattaaataa tctatacaaa cttttttaac acaaatgcat
     2761 acatatagcc agtgggattt gtagctagtc gatcgactct agaggatcag aaaattatcg
     2821 ccataaaaga cagaataagt catcagcggt tgtttcattt cctatatttt ttttttattt
     2881 ttttattttt taataaggga aaatttaacg tctaaggata cagaagattg ttagcacatt
     2941 aaagtaataa aggcttaagt agtaagtgcc ttagcatgtt attgtatttc aaaggacata
     3001 atctaaaata ataacaatat catttctcac aagttattca attttctttt ttttttctaa
     3061 taatatcaag aatgtattat ttgtttgaca taagtcaact aatttattta atatgctgga
     3121 ttaatcttgc agacatgtaa attaacaagt tttagtcaaa taacgttgaa gtttcaatga
     3181 actcaaataa tttctctttt tttttatata accataatct gatttatatt ttccgcaggg
     3241 atcaactgaa gttatgacat ttggattgga tcacttataa ccttggtcgc caaataatac
     3301 aaaaatcagc gttataaaac aaagaaggtt tttgttaaga aattaatcct ctttcttgat
     3361 aagaaagttg aaccgaaatt gcagatactg atatatgaaa ataataccca caattttggg
     3421 aatagcgcaa gcctcaattt aaacaatagg tgaggacaca tgataatgac ctcaatgatt
     3481 gttagaagaa aagagcctca ttacaaaatc gaaaaatgaa tggttgggta caagtttcca
     3541 aaacatggta aagtggactt tgcgtatgag acgtaaatag aaaaaaacac ttgttatatg
     3601 ttttctagaa ttattgttgt ctctttatgg ttggatgatg caaaatagta atttcggtta
     3661 gttgctgtaa aacaccacga gacaaataga tatggatatt tattaaatca ggaaaaacgt
     3721 aactctcggc tactggatgg ttcagtcacc caacgattac tggggagaga aaacagggca
     3781 aaagcaaagc ttaaaggaat ccgattgtca ttcggcaatg tgcagcgaaa ctaaaaaccg
     3841 gataatggac ctgttaatcg aaacattgaa gataaaggaa gaggaatcct ggcatatcat
     3901 caattgaata agttgaatta attatttcaa tctcattctc actttctgac ttatagtcgc
     3961 tttgttaaat catatgtcga ctctagagga tccccgggta aaaggaatgt ctcccttgcc
     4021 agtactgcta gggtttttct ttcaaactat ggaagcccat tcaagctgca tattacgatt
     4081 ttgtttttcg cttttagaaa gtggtttaga tgagataata gaaaaattct tgatctccga
     4141 caacgagtac ttttattttt tttgctaatc actttactca atattagctc gaaatcgtag
     4201 aaacgtagac gggtgcggga taccgagtgg tgtagttaag aatttttata aaccacgtgg
     4261 cccaaaaata tgaacccaaa acgtttatac atgagtatac tttaagaagg ctatacccct
     4321 tcgtgttaga tgtagtttta gctacccaac ccgagtctat gagcttgact tcagatgtag
     4381 aaggcattaa atcgttttga atattaatta aaaaacgatg aaaattaaat atttaaaagc
     4441 aatcatacgc tgaaaattta gtgctgtggc taatccttca acatggaaat gccataaaag
     4501 tgactttgac aaaaaaaaaa gtatatacag gtagtaaact catctacttc attgactttg
     4561 tttacagcat gtggaaggag gaatatttat tgctaaatcg tagtttaaca ttcaataagt
     4621 aatactattg aaattcgaca agattggccg catggatgaa aaagaggcat tttgctttgg
     4681 gagaattagt ccaaattaga actgaaaaaa aaactttacg aggcaaaaat gtcggattga
     4741 gatcgtaaaa gttcgctcgt cgtcttttgc tttgtgattg ttttcatgga tacatcttgc
     4801 tggatattta aattttagta ctatgtataa gatattctat aaatgtttta tcacccaaac
     4861 ctgttagcgc cttcttaatt ctattcaatc tggcttttgc tctgagacta cttcttggac
     4921 tttcactact tgttagttat acggaatttg tgtaattaga agtgaaataa tcctttctat
     4981 tagtaatgcg agctgcaatt cgagtctaac tccttaacca ctcggacata gtgacttatc
     5041 tgacactcat tgtaaattaa tatatatata ggcattttgt ttagttaaag gtacttaagt
     5101 aattagtata aacgaaccaa ttttataatc aggaagttaa gtgaatggta gcacatgtcg
     5161 taaaaattgt gaatttttat tgaataatat tttaaataca agcctttcta gactaggtat
     5221 actcataaac atatatgagc aaaaggatag aggagattac attgcatctt ctacaaattt
     5281 atttattgcc ctttactgaa aaattaaata atgagtacta aatgataaaa agcgctcagt
     5341 acaagaaaga ttgcaaaaat attgcattct tcatgaatta aagttgcata taaggcatat
     5401 tgaaagtaat agtactaaaa cagcagttag cgaaaattaa tagaattata ttcgcaagac
     5461 aattgtgaca ttaaattaaa aaattgtaga atttttacta tcctctttaa cgccatgagc
     5521 ctttataaaa aggttaaatt agttttaaca ttcttttttt gagtaagagt ttacaattta
     5581 tcaaaacctg tgttattata ttcattagtt tcaatttatt agcatctaga gaaaaatcaa
     5641 ttggcagtta cattgttgga atttatgaag aaaaagaatc tacaacggag aataagttgc
     5701 tgatcgcttt ccctaaaatt gtatattttg ctgagcttat tttgacattt cgttgaagtt
     5761 ttctctaatt cgcattcatt ttaagtaaaa caatgagaaa taaaattaca aaaaatacaa
     5821 attaaaatac aatttttagc tataattata gacgatgccc ttgtatccca ttctgtctcg
     5881 cttgccccta ctttttatct tttatatacc ataatgaacg ctgccgctac taaccatacc
     5941 ccgattttac atttcggact cccaaggacg tacaaaatag aaaactatag aaaaaaataa
     6001 tcagaaaata gcatgtcatc tctttgtaaa acgcgtttgc aagaagaaag gaaacaatgg
     6061 agaagagatc atccatttgt atgtaaattt tagtaaactt gaagaaatca ctaacaactt
     6121 ctcttactta gggattctat gcaaaacctt gtaaatcatc tgatggagga ctcgatttaa
     6181 tgaattggaa ggttggattt ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6241 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6301 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6361 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6421 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6481 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6541 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6601 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6661 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6721 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6781 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6841 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6901 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6961 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7021 ctgataaatc tggagccggt gagcgtggat ctcgcggtat cattgcagca ctggggccag
     7081 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7141 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7201 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7261 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7321 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7381 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7441 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7501 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7561 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7621 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7681 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7741 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7801 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7861 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7921 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7981 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8041 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     8101 agcgcagcga gtcagtgagc gaggaagcgg aag
//
