LOCUS       sequence-199301-        8563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt1 promoter with Sp.lys9 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8563)
  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 8563)
  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: pYZ174
FEATURES             Location/Qualifiers
     source          1..8563
                     /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"
     RBS             3112..3123
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        3243..4401
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     rep_origin      5499..6282
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6658..6762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6763..6831,6832..7623)
                     /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(6981..7000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7794..8382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8283..8302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8536..8553
                     /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 ttacaatttt gttcaaataa tcgagaagag gttatttttt cttcgagttt tacgtcggtt
     1081 acctaagact aaatacatct acttagaaaa tacgaaaacc ccaagtaaag cttcacataa
     1141 ataaggttgc aaaacaaact gaacatttat acttatgaac atgtagataa attgccaaaa
     1201 aaaaggttga cgaactctaa gattacgaat aattatagaa aatgaagaag ctttggtgtt
     1261 ctaaaatgtc aaacttatat ataagagttt ggaaccaaag cgtgaaacat gcttactctt
     1321 cgtcaataat ttcttcctcc aatcgaatac cttctttggc aagtgtatca atcaatggac
     1381 cgcaaagctt catatcatta ggagccaaaa cacctggggt gttaataaca ccatcaagaa
     1441 tttgctgagt agcaacacca caaggaacgc cgactaattt tgccatagag gtgtagccat
     1501 taggaacacc gtagtcaagc aaagtgcaag tacgagtttg gcgttttcct tctttggttt
     1561 caacttcaaa cttgtgttga aggattaaca tatcacgttc accttcttcg tattgcataa
     1621 gttcctccaa ggtagcgcat aaggtgtcca aaggattacc acgaggagta acgggtttgc
     1681 tagagaacat accaagccac ttcaaaccat tcaaaatgcg cttcttgtcg tcttcatctt
     1741 tgaatttgtg gatggaagaa attttcttga ttaggtcggc ttcggaggaa gattcagcct
     1801 tgatgacacg agcagtaacc tccttccaag gcaaagcggg tgcttcagga gataagtatt
     1861 cctgagctgt ctcatccaaa aatcccatat caaccaaaca atgaataaac tcgggaaatc
     1921 cttggtaacg taaagtaccg cgaatgatag tttctgcttc aggtatttga tagcgctctt
     1981 ggtagacagt ggaatcacga ttaggataac aaacgaaagc gtaaccggga tagatgaaat
     2041 agggtttagc agtttccatc aaatccttgc cgtcaatttc aacaagtttg ccgttttcat
     2101 aaaacttggc agagttgcga agagccaaca aaacaccacg agaagaccaa gaaaacttgt
     2161 atcctaaagg attgttggag tcctcgggag cgggaagacc tccacaataa gacaagaaag
     2221 acttaatctt accgccagct ttatgaacct cttcaatggt cttgatagca tataaatgat
     2281 caataccggg atcgacgccg atttcgttca tacaaataga accagccttg atggcagctt
     2341 cctccaactc agccattttg ggattcacat aactggtggt acaaacgtgt ttaccatgct
     2401 taatggcagc tttcataact gttgcatgat aagtgtaagg aatcaagctg atggtaagat
     2461 catgttcgga aacagctttc tcaagagctg cttcatcatt gacatccaaa gcaatggctt
     2521 tgctgtttgg gataccatta ataaatgctt cagccttaga aagagttctg catgctacgg
     2581 ttatattatt ttctttacga cgagataaat attctaaagt aggatgagcg acaaatcccg
     2641 aaccaagtaa taagatggaa ggcattttga tgttgaaaat gaaagaggca gtcagttaag
     2701 gacaagtggt aaaccaaaga cttctgtttt gtcgtcacgg ttttagcgat atataagggg
     2761 ttccttttta tccacagatt tatttctgat ggccgaaatc gtcgtctaaa ttgatgaaaa
     2821 ctgattatct cgaatcggtt aaaaaagcag tcacatggac cattttttga attaatattt
     2881 aatcgttacg tttcctctca atgttatgtt tggcttacta ggggaattgg ttattatcga
     2941 caagtttagt cgataatttg ttagcagcga tgtcgcaact gatagtttta gcaacttttt
     3001 ttacgtagtt tttttctgca caacagaagt tgcttaaaag ctagtagatg ccattcgttg
     3061 ttgtttgggg gtagattaaa gaattgtaag ctttagtata agaataattt gtttctcctc
     3121 tttctgccta tctgttgttc ataatcaact gcgtagttct ggtgaaggag taaattaaac
     3181 tgcctcgcac cgccgtactc atttttacct cggtcgatcg actctagagg atcagaaaat
     3241 tatcgccata aaagacagaa taagtcatca gcggttgttt catttcctat attttttttt
     3301 tattttttta ttttttaata agggaaaatt taacgtctaa ggatacagaa gattgttagc
     3361 acattaaagt aataaaggct taagtagtaa gtgccttagc atgttattgt atttcaaagg
     3421 acataatcta aaataataac aatatcattt ctcacaagtt attcaatttt cttttttttt
     3481 tctaataata tcaagaatgt attatttgtt tgacataagt caactaattt atttaatatg
     3541 ctggattaat cttgcagaca tgtaaattaa caagttttag tcaaataacg ttgaagtttc
     3601 aatgaactca aataatttct cttttttttt atataaccat aatctgattt atattttccg
     3661 cagggatcaa ctgaagttat gacatttgga ttggatcact tataaccttg gtcgccaaat
     3721 aatacaaaaa tcagcgttat aaaacaaaga aggtttttgt taagaaatta atcctctttc
     3781 ttgataagaa agttgaaccg aaattgcaga tactgatata tgaaaataat acccacaatt
     3841 ttgggaatag cgcaagcctc aatttaaaca ataggtgagg acacatgata atgacctcaa
     3901 tgattgttag aagaaaagag cctcattaca aaatcgaaaa atgaatggtt gggtacaagt
     3961 ttccaaaaca tggtaaagtg gactttgcgt atgagacgta aatagaaaaa aacacttgtt
     4021 atatgttttc tagaattatt gttgtctctt tatggttgga tgatgcaaaa tagtaatttc
     4081 ggttagttgc tgtaaaacac cacgagacaa atagatatgg atatttatta aatcaggaaa
     4141 aacgtaactc tcggctactg gatggttcag tcacccaacg attactgggg agagaaaaca
     4201 gggcaaaagc aaagcttaaa ggaatccgat tgtcattcgg caatgtgcag cgaaactaaa
     4261 aaccggataa tggacctgtt aatcgaaaca ttgaagatat ataaaggaag aggaatcctg
     4321 gcatatcatc aattgaataa gttgaattaa ttatttcaat ctcattctca ctttctgact
     4381 tatagtcgct ttgttaaatc atatgtcgac tctagaggat ccccgggtaa aaggaatgtc
     4441 tcccttgcca gtactgctag ggtttttctt tcaaactatg gaagcccatt caagctgcat
     4501 attacgattt tgtttttcgc ttttagaaag tggtttagat gagataatag aaaaattctt
     4561 gatctccgac aacgagtact tttatttttt ttgctaatca ctttactcaa tattagctcg
     4621 aaatcgtaga aacgtagacg ggtgcgggat accgagtggt gtagttaaga atttttataa
     4681 accacgtggc ccaaaaatat gaacccaaaa cgtttataca tgagtatact ttaagaaggc
     4741 tatacccctt cgtgttagat gtagttttag ctacccaacc cgagtctatg agcttgactt
     4801 cagatgtaga aggcattaaa tcgttttgaa tattaattaa aaaacgatga aaattaaata
     4861 tttaaaagca atcatacgct gaaaatttag tgctgtggct aatccttcaa catggaaatg
     4921 ccataaaagt gactttgaca aaaaaaaaag tatatacagg tagtaaactc atctacttca
     4981 ttgactttgt ttacagcatg tggaaggagg aatatttatt gctaaatcgt agtttaacat
     5041 tcaataagta atactattga aattcgacaa gattggccgc atggatgaaa aagaggcatt
     5101 ttgctttggg agaattagtc caaattagaa ctgaaaaaaa aaactttacg aggcaaaaat
     5161 gtcggattga gatcgtaaaa gttcgctcgt cgtcttttgc tttgtgattg ttttcatgga
     5221 tacatcttgc tggatattta aattttagta ctatgtataa gatattctat aaatgtttta
     5281 tcacccaaac ctgttagcgc cttcttaatt ctattcaatc tggcttttgc tctgagacta
     5341 cttcttggac tttcactact tgttagttat acggaatttg tgtaattaga agtgaaataa
     5401 tcctttctat tagtaatgcg agctgcaatt cgagtctaac tccttaacca ctcggacata
     5461 gtgacttatc tgacactcat tgtaaattaa tatatatata ggcattttgt ttagttaaag
     5521 gtacttaagt aattagtata aacgaaccaa ttttataatc aggaagttaa gtgaatggta
     5581 gcacatgtcg taaaaattgt gaatttttat tgaataatat tttaaataca agcctttcta
     5641 gactaggtat actcataaac atatatgagc aaaaggatag aggagattac attgcatctt
     5701 ctacaaattt atttattgcc ctttactgaa aaattaaata atgagtacta aatgataaaa
     5761 agcgctcagt acaagaaaga ttgcaaaaat attgcattct tcatgaatta aagttgcata
     5821 taaggcatat tgaaagtaat agtactaaaa cagcagttag cgaaaattaa tagaattata
     5881 ttcgcaagac aattgtgaca ttaaattaaa aaattgtaga atttttacta tcctctttaa
     5941 cgccatgagc ctttataaaa aggttaaatt agttttaaca ttcttttttt gagtaagagt
     6001 ttacaattta tcaaaacctg tgttattata ttcattagtt tcaatttatt agcatctaga
     6061 gaaaaatcaa ttggcagtta cattgttgga atttatgaag aaaaagaatc tacaacggag
     6121 aataagttgc tgatcgcttt ccctaaaatt gtatattttg ctgagcttat tttgacattt
     6181 cgttgaagtt ttctctaatt cgcattcatt ttaagtaaaa caatgagaaa taaaattaca
     6241 aaaaatacaa attaaaatac aatttttagc tataattata gacgatgccc ttgtatccca
     6301 ttctgtctcg cttgccccta ctttttatct tttatatacc ataatgaacg ctgccgctac
     6361 taaccatacc ccgattttac atttcggact cccaaggacg tacaaaatag aaaactatag
     6421 aaaaaaataa tcagaaaata gcatgtcatc tctttgtaaa acgcgtttgc aagaagaaag
     6481 gaaacaatgg agaagagatc atccatttgt atgtaaattt tagtaaactt gaagaaatca
     6541 ctaacaactt ctcttactta gggattctat gcaaaacctt gtaaatcatc tgatggagga
     6601 ctcgatttaa tgaattggaa ggttggattt ccaggtggca cttttcgggg aaatgtgcgc
     6661 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6721 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6781 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6841 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6901 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6961 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7021 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7081 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7141 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7201 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7261 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7321 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7381 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7441 tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat cattgcagca
     7501 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7561 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7621 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7681 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7741 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7861 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7921 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7981 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8041 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8101 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8161 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8221 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8281 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8341 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8401 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8461 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8521 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
