LOCUS       sequence-198902-        8138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt1 promoter with Sp.ura4 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8138)
  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 8138)
  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: pYZ155
FEATURES             Location/Qualifiers
     source          1..8138
                     /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..3976
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     rep_origin      5074..5857
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6233..6337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6338..6406,6407..7198)
                     /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(6556..6575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7369..7957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7858..7877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8111..8128
                     /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 gatatataaa ggaagaggaa tcctggcata
     3901 tcatcaattg aataagttga attaattatt tcaatctcat tctcactttc tgacttatag
     3961 tcgctttgtt aaatcatatg tcgactctag aggatccccg ggtaaaagga atgtctccct
     4021 tgccagtact gctagggttt ttctttcaaa ctatggaagc ccattcaagc tgcatattac
     4081 gattttgttt ttcgctttta gaaagtggtt tagatgagat aatagaaaaa ttcttgatct
     4141 ccgacaacga gtacttttat tttttttgct aatcacttta ctcaatatta gctcgaaatc
     4201 gtagaaacgt agacgggtgc gggataccga gtggtgtagt taagaatttt tataaaccac
     4261 gtggcccaaa aatatgaacc caaaacgttt atacatgagt atactttaag aaggctatac
     4321 cccttcgtgt tagatgtagt tttagctacc caacccgagt ctatgagctt gacttcagat
     4381 gtagaaggca ttaaatcgtt ttgaatatta attaaaaaac gatgaaaatt aaatatttaa
     4441 aagcaatcat acgctgaaaa tttagtgctg tggctaatcc ttcaacatgg aaatgccata
     4501 aaagtgactt tgacaaaaaa aaaagtatat acaggtagta aactcatcta cttcattgac
     4561 tttgtttaca gcatgtggaa ggaggaatat ttattgctaa atcgtagttt aacattcaat
     4621 aagtaatact attgaaattc gacaagattg gccgcatgga tgaaaaagag gcattttgct
     4681 ttgggagaat tagtccaaat tagaactgaa aaaaaaaact ttacgaggca aaaatgtcgg
     4741 attgagatcg taaaagttcg ctcgtcgtct tttgctttgt gattgttttc atggatacat
     4801 cttgctggat atttaaattt tagtactatg tataagatat tctataaatg ttttatcacc
     4861 caaacctgtt agcgccttct taattctatt caatctggct tttgctctga gactacttct
     4921 tggactttca ctacttgtta gttatacgga atttgtgtaa ttagaagtga aataatcctt
     4981 tctattagta atgcgagctg caattcgagt ctaactcctt aaccactcgg acatagtgac
     5041 ttatctgaca ctcattgtaa attaatatat atataggcat tttgtttagt taaaggtact
     5101 taagtaatta gtataaacga accaatttta taatcaggaa gttaagtgaa tggtagcaca
     5161 tgtcgtaaaa attgtgaatt tttattgaat aatattttaa atacaagcct ttctagacta
     5221 ggtatactca taaacatata tgagcaaaag gatagaggag attacattgc atcttctaca
     5281 aatttattta ttgcccttta ctgaaaaatt aaataatgag tactaaatga taaaaagcgc
     5341 tcagtacaag aaagattgca aaaatattgc attcttcatg aattaaagtt gcatataagg
     5401 catattgaaa gtaatagtac taaaacagca gttagcgaaa attaatagaa ttatattcgc
     5461 aagacaattg tgacattaaa ttaaaaaatt gtagaatttt tactatcctc tttaacgcca
     5521 tgagccttta taaaaaggtt aaattagttt taacattctt tttttgagta agagtttaca
     5581 atttatcaaa acctgtgtta ttatattcat tagtttcaat ttattagcat ctagagaaaa
     5641 atcaattggc agttacattg ttggaattta tgaagaaaaa gaatctacaa cggagaataa
     5701 gttgctgatc gctttcccta aaattgtata ttttgctgag cttattttga catttcgttg
     5761 aagttttctc taattcgcat tcattttaag taaaacaatg agaaataaaa ttacaaaaaa
     5821 tacaaattaa aatacaattt ttagctataa ttatagacga tgcccttgta tcccattctg
     5881 tctcgcttgc ccctactttt tatcttttat ataccataat gaacgctgcc gctactaacc
     5941 ataccccgat tttacatttc ggactcccaa ggacgtacaa aatagaaaac tatagaaaaa
     6001 aataatcaga aaatagcatg tcatctcttt gtaaaacgcg tttgcaagaa gaaaggaaac
     6061 aatggagaag agatcatcca tttgtatgta aattttagta aacttgaaga aatcactaac
     6121 aacttctctt acttagggat tctatgcaaa accttgtaaa tcatctgatg gaggactcga
     6181 tttaatgaat tggaaggttg gatttccagg tggcactttt cggggaaatg tgcgcggaac
     6241 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6301 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6361 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6421 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6481 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6541 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6601 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6661 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6721 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6781 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6841 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6901 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6961 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7021 tattgctgat aaatctggag ccggtgagcg tggatctcgc ggtatcattg cagcactggg
     7081 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7141 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7201 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7261 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7321 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7381 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7441 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7501 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7561 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7621 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7681 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7741 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7801 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7861 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7921 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7981 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8041 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8101 gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
