LOCUS       sequence-198980-        8275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt1 promoter with Sp.leu1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8275)
  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 8275)
  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: pYZ158
FEATURES             Location/Qualifiers
     source          1..8275
                     /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"
     promoter        2955..4113
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     rep_origin      5211..5994
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6370..6474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6475..6543,6544..7335)
                     /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(6693..6712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7506..8094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7995..8014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8248..8265
                     /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 ttacaatttc aactcggtca taaagttgaa cggatgtcgt aaatcaattc catgcttttg
     1081 caacttaata catctttgct tgactataac ataccaaaag aaaaaacatt agagccttaa
     1141 aatttacgct tttgtcaatt gcatttttaa cgataataaa tatgcgacac aattaatcaa
     1201 taaaaattag gtcaaatcaa atgatatgaa tttattctac attaaaccct attacaaaat
     1261 tttttcaagt tcttcaacaa cagccttagt aatatcagcg gtagaagcct cacctcccaa
     1321 atcgcgagta taaagaccac gtccaccaat tgaagtatca tccaagacct tgcgtacggc
     1381 ggcttcgata gcttcagcct ccttaggagc attcaaacca taacgaagga gaagggaagc
     1441 agataaaatt gtaccaacag gattaacaat gcccttgcca gcgatatcgg gagcgctacc
     1501 gtgaatgggc tcaaccaaac aatgaacctt ttcttctgat tttcctacca caccggaaag
     1561 ggaggcagaa ggcaaaaggc ccaagctacc aggaatgaca gaagcctcat ctgaaataat
     1621 gtcaccaaac aagttgtcag tcaaaacaac accgttaagt gtacgagggc tcttgaccaa
     1681 aagcatggct gcggagtcaa tgagctggtt ttttaaggta aggtgaggat attcctcctt
     1741 aaaaatctta gctacagtct tgcgccaaag acgagaagtt gccaaaacat tagctttgtc
     1801 gagtaatgtg acgggagcag gagggttgga agtttcagct aaccaagcag ccaaacgagc
     1861 aatacgagaa acttcttcca aactgtaagg ccaagtgtcc atagcataac ccgatccgtt
     1921 gtcctcagtg cgctcaccaa agtaacaacc tccagtaagt tctcgtacaa cacaaaaatc
     1981 gacaccttca acgatttcag gcttcaaagg gctgtacttg actaaagact tgctggcaaa
     2041 gttgcaaggt cgaaggttgg cccaaacacc catactctta cgaagcttca ataaaccttg
     2101 ctcaggacga caattggggt tggtccattc aggaccacca acggcaccca aaagaacacc
     2161 gtcagcttcc aaacaagcct tcacagtctc gtcagtcaaa ggggttccat aggcatcaat
     2221 agaggcacct ccaatcttgt gttcttcaaa ctccagtttt aactcaggtc gcttcttctc
     2281 aacgactttc aaaacctcca aggcagaagc aacaatttca gggccaatat gatctcctgg
     2341 taagacgacg attttctttg cacacatgtt gttgaagaag ttttgttgtg aaatggtttc
     2401 gtgaaagttt cagaccctac cgcaaaaatg cctggtttcg ggaaactcaa cactgttgca
     2461 ctttttatac tacagattgg gatatcgata atattgcgta aaaaatcctt tttttaaaaa
     2521 gcttgtttac agtaacgtaa atgaccagaa atcagatgaa aatcacaaga aagcaaataa
     2581 ttcacgttaa atcctgatat gtttgatttt gtgatgaaat catggatgtt cataggaatt
     2641 gttgaaattg cgctttttta acgaaatata caagtatcct ggagcttact taattaatta
     2701 atgaatcttt gtttctagat attaaaatag tagcctcaat tatcagcgct ttctacgtta
     2761 gtaaacgaaa tttttaatgt caaaaaaatg tttaatagac agtacgaata tgcattataa
     2821 tgttcataaa taatttgagt atgtgtacga gttgtcttta aacccacaga ggtagaatgt
     2881 atatataaaa ttaataagct aagtgtaata cttaaaaaat acatgtcgat cgactctaga
     2941 ggatcagaaa attatcgcca taaaagacag aataagtcat cagcggttgt ttcatttcct
     3001 atattttttt tttatttttt tattttttaa taagggaaaa tttaacgtct aaggatacag
     3061 aagattgtta gcacattaaa gtaataaagg cttaagtagt aagtgcctta gcatgttatt
     3121 gtatttcaaa ggacataatc taaaataata acaatatcat ttctcacaag ttattcaatt
     3181 ttcttttttt tttctaataa tatcaagaat gtattatttg tttgacataa gtcaactaat
     3241 ttatttaata tgctggatta atcttgcaga catgtaaatt aacaagtttt agtcaaataa
     3301 cgttgaagtt tcaatgaact caaataattt ctcttttttt ttatataacc ataatctgat
     3361 ttatattttc cgcagggatc aactgaagtt atgacatttg gattggatca cttataacct
     3421 tggtcgccaa ataatacaaa aatcagcgtt ataaaacaaa gaaggttttt gttaagaaat
     3481 taatcctctt tcttgataag aaagttgaac cgaaattgca gatactgata tatgaaaata
     3541 atacccacaa ttttgggaat agcgcaagcc tcaatttaaa caataggtga ggacacatga
     3601 taatgacctc aatgattgtt agaagaaaag agcctcatta caaaatcgaa aaatgaatgg
     3661 ttgggtacaa gtttccaaaa catggtaaag tggactttgc gtatgagacg taaatagaaa
     3721 aaaacacttg ttatatgttt tctagaatta ttgttgtctc tttatggttg gatgatgcaa
     3781 aatagtaatt tcggttagtt gctgtaaaac accacgagac aaatagatat ggatatttat
     3841 taaatcagga aaaacgtaac tctcggctac tggatggttc agtcacccaa cgattactgg
     3901 ggagagaaaa cagggcaaaa gcaaagctta aaggaatccg attgtcattc ggcaatgtgc
     3961 agcgaaacta aaaaccggat aatggacctg ttaatcgaaa cattgaagat atataaagga
     4021 agaggaatcc tggcatatca tcaattgaat aagttgaatt aattatttca atctcattct
     4081 cactttctga cttatagtcg ctttgttaaa tcatatgtcg actctagagg atccccgggt
     4141 aaaaggaatg tctcccttgc cagtactgct agggtttttc tttcaaacta tggaagccca
     4201 ttcaagctgc atattacgat tttgtttttc gcttttagaa agtggtttag atgagataat
     4261 agaaaaattc ttgatctccg acaacgagta cttttatttt ttttgctaat cactttactc
     4321 aatattagct cgaaatcgta gaaacgtaga cgggtgcggg ataccgagtg gtgtagttaa
     4381 gaatttttat aaaccacgtg gcccaaaaat atgaacccaa aacgtttata catgagtata
     4441 ctttaagaag gctatacccc ttcgtgttag atgtagtttt agctacccaa cccgagtcta
     4501 tgagcttgac ttcagatgta gaaggcatta aatcgttttg aatattaatt aaaaaacgat
     4561 gaaaattaaa tatttaaaag caatcatacg ctgaaaattt agtgctgtgg ctaatccttc
     4621 aacatggaaa tgccataaaa gtgactttga caaaaaaaaa agtatataca ggtagtaaac
     4681 tcatctactt cattgacttt gtttacagca tgtggaagga ggaatattta ttgctaaatc
     4741 gtagtttaac attcaataag taatactatt gaaattcgac aagattggcc gcatggatga
     4801 aaaagaggca ttttgctttg ggagaattag tccaaattag aactgaaaaa aaaaacttta
     4861 cgaggcaaaa atgtcggatt gagatcgtaa aagttcgctc gtcgtctttt gctttgtgat
     4921 tgttttcatg gatacatctt gctggatatt taaattttag tactatgtat aagatattct
     4981 ataaatgttt tatcacccaa acctgttagc gccttcttaa ttctattcaa tctggctttt
     5041 gctctgagac tacttcttgg actttcacta cttgttagtt atacggaatt tgtgtaatta
     5101 gaagtgaaat aatcctttct attagtaatg cgagctgcaa ttcgagtcta actccttaac
     5161 cactcggaca tagtgactta tctgacactc attgtaaatt aatatatata taggcatttt
     5221 gtttagttaa aggtacttaa gtaattagta taaacgaacc aattttataa tcaggaagtt
     5281 aagtgaatgg tagcacatgt cgtaaaaatt gtgaattttt attgaataat attttaaata
     5341 caagcctttc tagactaggt atactcataa acatatatga gcaaaaggat agaggagatt
     5401 acattgcatc ttctacaaat ttatttattg ccctttactg aaaaattaaa taatgagtac
     5461 taaatgataa aaagcgctca gtacaagaaa gattgcaaaa atattgcatt cttcatgaat
     5521 taaagttgca tataaggcat attgaaagta atagtactaa aacagcagtt agcgaaaatt
     5581 aatagaatta tattcgcaag acaattgtga cattaaatta aaaaattgta gaatttttac
     5641 tatcctcttt aacgccatga gcctttataa aaaggttaaa ttagttttaa cattcttttt
     5701 ttgagtaaga gtttacaatt tatcaaaacc tgtgttatta tattcattag tttcaattta
     5761 ttagcatcta gagaaaaatc aattggcagt tacattgttg gaatttatga agaaaaagaa
     5821 tctacaacgg agaataagtt gctgatcgct ttccctaaaa ttgtatattt tgctgagctt
     5881 attttgacat ttcgttgaag ttttctctaa ttcgcattca ttttaagtaa aacaatgaga
     5941 aataaaatta caaaaaatac aaattaaaat acaattttta gctataatta tagacgatgc
     6001 ccttgtatcc cattctgtct cgcttgcccc tactttttat cttttatata ccataatgaa
     6061 cgctgccgct actaaccata ccccgatttt acatttcgga ctcccaagga cgtacaaaat
     6121 agaaaactat agaaaaaaat aatcagaaaa tagcatgtca tctctttgta aaacgcgttt
     6181 gcaagaagaa aggaaacaat ggagaagaga tcatccattt gtatgtaaat tttagtaaac
     6241 ttgaagaaat cactaacaac ttctcttact tagggattct atgcaaaacc ttgtaaatca
     6301 tctgatggag gactcgattt aatgaattgg aaggttggat ttccaggtgg cacttttcgg
     6361 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6421 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6481 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6541 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6601 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6661 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6721 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6781 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6841 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6901 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6961 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7021 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7081 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7141 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg atctcgcggt
     7201 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7261 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7321 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7381 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7441 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7501 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7561 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7621 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7681 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7741 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7801 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7861 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7921 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7981 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8041 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8101 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8161 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8221 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
