LOCUS       sequence-199305-        8558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt41 promoter with Sp.lys9 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8558)
  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 8558)
  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: pYZ175
FEATURES             Location/Qualifiers
     source          1..8558
                     /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..4397
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      5494..6277
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6653..6757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6758..6826,6827..7618)
                     /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(6976..6995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7789..8377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8278..8297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8531..8548
                     /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 ttgaagataa aggaagagga atcctggcat
     4321 atcatcaatt gaataagttg aattaattat ttcaatctca ttctcacttt ctgacttata
     4381 gtcgctttgt taaatcatat gtcgactcta gaggatcccc gggtaaaagg aatgtctccc
     4441 ttgccagtac tgctagggtt tttctttcaa actatggaag cccattcaag ctgcatatta
     4501 cgattttgtt tttcgctttt agaaagtggt ttagatgaga taatagaaaa attcttgatc
     4561 tccgacaacg agtactttta ttttttttgc taatcacttt actcaatatt agctcgaaat
     4621 cgtagaaacg tagacgggtg cgggataccg agtggtgtag ttaagaattt ttataaacca
     4681 cgtggcccaa aaatatgaac ccaaaacgtt tatacatgag tatactttaa gaaggctata
     4741 ccccttcgtg ttagatgtag ttttagctac ccaacccgag tctatgagct tgacttcaga
     4801 tgtagaaggc attaaatcgt tttgaatatt aattaaaaaa cgatgaaaat taaatattta
     4861 aaagcaatca tacgctgaaa atttagtgct gtggctaatc cttcaacatg gaaatgccat
     4921 aaaagtgact ttgacaaaaa aaaaagtata tacaggtagt aaactcatct acttcattga
     4981 ctttgtttac agcatgtgga aggaggaata tttattgcta aatcgtagtt taacattcaa
     5041 taagtaatac tattgaaatt cgacaagatt ggccgcatgg atgaaaaaga ggcattttgc
     5101 tttgggagaa ttagtccaaa ttagaactga aaaaaaaact ttacgaggca aaaatgtcgg
     5161 attgagatcg taaaagttcg ctcgtcgtct tttgctttgt gattgttttc atggatacat
     5221 cttgctggat atttaaattt tagtactatg tataagatat tctataaatg ttttatcacc
     5281 caaacctgtt agcgccttct taattctatt caatctggct tttgctctga gactacttct
     5341 tggactttca ctacttgtta gttatacgga atttgtgtaa ttagaagtga aataatcctt
     5401 tctattagta atgcgagctg caattcgagt ctaactcctt aaccactcgg acatagtgac
     5461 ttatctgaca ctcattgtaa attaatatat atataggcat tttgtttagt taaaggtact
     5521 taagtaatta gtataaacga accaatttta taatcaggaa gttaagtgaa tggtagcaca
     5581 tgtcgtaaaa attgtgaatt tttattgaat aatattttaa atacaagcct ttctagacta
     5641 ggtatactca taaacatata tgagcaaaag gatagaggag attacattgc atcttctaca
     5701 aatttattta ttgcccttta ctgaaaaatt aaataatgag tactaaatga taaaaagcgc
     5761 tcagtacaag aaagattgca aaaatattgc attcttcatg aattaaagtt gcatataagg
     5821 catattgaaa gtaatagtac taaaacagca gttagcgaaa attaatagaa ttatattcgc
     5881 aagacaattg tgacattaaa ttaaaaaatt gtagaatttt tactatcctc tttaacgcca
     5941 tgagccttta taaaaaggtt aaattagttt taacattctt tttttgagta agagtttaca
     6001 atttatcaaa acctgtgtta ttatattcat tagtttcaat ttattagcat ctagagaaaa
     6061 atcaattggc agttacattg ttggaattta tgaagaaaaa gaatctacaa cggagaataa
     6121 gttgctgatc gctttcccta aaattgtata ttttgctgag cttattttga catttcgttg
     6181 aagttttctc taattcgcat tcattttaag taaaacaatg agaaataaaa ttacaaaaaa
     6241 tacaaattaa aatacaattt ttagctataa ttatagacga tgcccttgta tcccattctg
     6301 tctcgcttgc ccctactttt tatcttttat ataccataat gaacgctgcc gctactaacc
     6361 ataccccgat tttacatttc ggactcccaa ggacgtacaa aatagaaaac tatagaaaaa
     6421 aataatcaga aaatagcatg tcatctcttt gtaaaacgcg tttgcaagaa gaaaggaaac
     6481 aatggagaag agatcatcca tttgtatgta aattttagta aacttgaaga aatcactaac
     6541 aacttctctt acttagggat tctatgcaaa accttgtaaa tcatctgatg gaggactcga
     6601 tttaatgaat tggaaggttg gatttccagg tggcactttt cggggaaatg tgcgcggaac
     6661 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6721 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6781 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6841 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6901 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6961 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7021 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7081 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7141 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7201 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7261 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7321 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7381 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7441 tattgctgat aaatctggag ccggtgagcg tggatctcgc ggtatcattg cagcactggg
     7501 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7561 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7621 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7681 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7741 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7801 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7861 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7921 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7981 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8041 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     8101 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8161 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     8221 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8281 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8341 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8401 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8461 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8521 gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
