LOCUS       sequence-199307-        8556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt81 promoter with Sp.his3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8556)
  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 8556)
  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: pYZ176
FEATURES             Location/Qualifiers
     source          1..8556
                     /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..4394
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     rep_origin      5492..6275
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6651..6755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6756..6824,6825..7616)
                     /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(6974..6993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7787..8375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8276..8295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8529..8546
                     /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 ttgaagatgg aagaggaatc ctggcatatc
     4321 atcaattgaa taagttgaat taattatttc aatctcattc tcactttctg acttatagtc
     4381 gctttgttaa atcatatgtc gactctagag gatccccggg taaaaggaat gtctcccttg
     4441 ccagtactgc tagggttttt ctttcaaact atggaagccc attcaagctg catattacga
     4501 ttttgttttt cgcttttaga aagtggttta gatgagataa tagaaaaatt cttgatctcc
     4561 gacaacgagt acttttattt tttttgctaa tcactttact caatattagc tcgaaatcgt
     4621 agaaacgtag acgggtgcgg gataccgagt ggtgtagtta agaattttta taaaccacgt
     4681 ggcccaaaaa tatgaaccca aaacgtttat acatgagtat actttaagaa ggctataccc
     4741 cttcgtgtta gatgtagttt tagctaccca acccgagtct atgagcttga cttcagatgt
     4801 agaaggcatt aaatcgtttt gaatattaat taaaaaacga tgaaaattaa atatttaaaa
     4861 gcaatcatac gctgaaaatt tagtgctgtg gctaatcctt caacatggaa atgccataaa
     4921 agtgactttg acaaaaaaaa aagtatatac aggtagtaaa ctcatctact tcattgactt
     4981 tgtttacagc atgtggaagg aggaatattt attgctaaat cgtagtttaa cattcaataa
     5041 gtaatactat tgaaattcga caagattggc cgcatggatg aaaaagaggc attttgcttt
     5101 gggagaatta gtccaaatta gaactgaaaa aaaaaacttt acgaggcaaa aatgtcggat
     5161 tgagatcgta aaagttcgct cgtcgtcttt tgctttgtga ttgttttcat ggatacatct
     5221 tgctggatat ttaaatttta gtactatgta taagatattc tataaatgtt ttatcaccca
     5281 aacctgttag cgccttctta attctattca atctggcttt tgctctgaga ctacttcttg
     5341 gactttcact acttgttagt tatacggaat ttgtgtaatt agaagtgaaa taatcctttc
     5401 tattagtaat gcgagctgca attcgagtct aactccttaa ccactcggac atagtgactt
     5461 atctgacact cattgtaaat taatatatat ataggcattt tgtttagtta aaggtactta
     5521 agtaattagt ataaacgaac caattttata atcaggaagt taagtgaatg gtagcacatg
     5581 tcgtaaaaat tgtgaatttt tattgaataa tattttaaat acaagccttt ctagactagg
     5641 tatactcata aacatatatg agcaaaagga tagaggagat tacattgcat cttctacaaa
     5701 tttatttatt gccctttact gaaaaattaa ataatgagta ctaaatgata aaaagcgctc
     5761 agtacaagaa agattgcaaa aatattgcat tcttcatgaa ttaaagttgc atataaggca
     5821 tattgaaagt aatagtacta aaacagcagt tagcgaaaat taatagaatt atattcgcaa
     5881 gacaattgtg acattaaatt aaaaaattgt agaattttta ctatcctctt taacgccatg
     5941 agcctttata aaaaggttaa attagtttta acattctttt tttgagtaag agtttacaat
     6001 ttatcaaaac ctgtgttatt atattcatta gtttcaattt attagcatct agagaaaaat
     6061 caattggcag ttacattgtt ggaatttatg aagaaaaaga atctacaacg gagaataagt
     6121 tgctgatcgc tttccctaaa attgtatatt ttgctgagct tattttgaca tttcgttgaa
     6181 gttttctcta attcgcattc attttaagta aaacaatgag aaataaaatt acaaaaaata
     6241 caaattaaaa tacaattttt agctataatt atagacgatg cccttgtatc ccattctgtc
     6301 tcgcttgccc ctacttttta tcttttatat accataatga acgctgccgc tactaaccat
     6361 accccgattt tacatttcgg actcccaagg acgtacaaaa tagaaaacta tagaaaaaaa
     6421 taatcagaaa atagcatgtc atctctttgt aaaacgcgtt tgcaagaaga aaggaaacaa
     6481 tggagaagag atcatccatt tgtatgtaaa ttttagtaaa cttgaagaaa tcactaacaa
     6541 cttctcttac ttagggattc tatgcaaaac cttgtaaatc atctgatgga ggactcgatt
     6601 taatgaattg gaaggttgga tttccaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6661 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6721 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6781 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6841 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6901 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6961 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7021 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7081 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7141 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7201 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7261 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7321 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7381 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7441 ttgctgataa atctggagcc ggtgagcgtg gatctcgcgg tatcattgca gcactggggc
     7501 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7561 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7621 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7681 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7741 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7801 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7861 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7921 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7981 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8041 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8101 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8161 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8221 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8281 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8341 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8401 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8461 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8521 ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
