LOCUS       sequence-205839-        8270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt41 promoter with Sp.leu1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8270)
  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 8270)
  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: pYZ159
FEATURES             Location/Qualifiers
     source          1..8270
                     /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..4109
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      5206..5989
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6365..6469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6470..6538,6539..7330)
                     /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(6688..6707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7501..8089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7990..8009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8243..8260
                     /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 aaaggaagag
     4021 gaatcctggc atatcatcaa ttgaataagt tgaattaatt atttcaatct cattctcact
     4081 ttctgactta tagtcgcttt gttaaatcat atgtcgactc tagaggatcc ccgggtaaaa
     4141 ggaatgtctc ccttgccagt actgctaggg tttttctttc aaactatgga agcccattca
     4201 agctgcatat tacgattttg tttttcgctt ttagaaagtg gtttagatga gataatagaa
     4261 aaattcttga tctccgacaa cgagtacttt tatttttttt gctaatcact ttactcaata
     4321 ttagctcgaa atcgtagaaa cgtagacggg tgcgggatac cgagtggtgt agttaagaat
     4381 ttttataaac cacgtggccc aaaaatatga acccaaaacg tttatacatg agtatacttt
     4441 aagaaggcta taccccttcg tgttagatgt agttttagct acccaacccg agtctatgag
     4501 cttgacttca gatgtagaag gcattaaatc gttttgaata ttaattaaaa aacgatgaaa
     4561 attaaatatt taaaagcaat catacgctga aaatttagtg ctgtggctaa tccttcaaca
     4621 tggaaatgcc ataaaagtga ctttgacaaa aaaaaaagta tatacaggta gtaaactcat
     4681 ctacttcatt gactttgttt acagcatgtg gaaggaggaa tatttattgc taaatcgtag
     4741 tttaacattc aataagtaat actattgaaa ttcgacaaga ttggccgcat ggatgaaaaa
     4801 gaggcatttt gctttgggag aattagtcca aattagaact gaaaaaaaaa ctttacgagg
     4861 caaaaatgtc ggattgagat cgtaaaagtt cgctcgtcgt cttttgcttt gtgattgttt
     4921 tcatggatac atcttgctgg atatttaaat tttagtacta tgtataagat attctataaa
     4981 tgttttatca cccaaacctg ttagcgcctt cttaattcta ttcaatctgg cttttgctct
     5041 gagactactt cttggacttt cactacttgt tagttatacg gaatttgtgt aattagaagt
     5101 gaaataatcc tttctattag taatgcgagc tgcaattcga gtctaactcc ttaaccactc
     5161 ggacatagtg acttatctga cactcattgt aaattaatat atatataggc attttgttta
     5221 gttaaaggta cttaagtaat tagtataaac gaaccaattt tataatcagg aagttaagtg
     5281 aatggtagca catgtcgtaa aaattgtgaa tttttattga ataatatttt aaatacaagc
     5341 ctttctagac taggtatact cataaacata tatgagcaaa aggatagagg agattacatt
     5401 gcatcttcta caaatttatt tattgccctt tactgaaaaa ttaaataatg agtactaaat
     5461 gataaaaagc gctcagtaca agaaagattg caaaaatatt gcattcttca tgaattaaag
     5521 ttgcatataa ggcatattga aagtaatagt actaaaacag cagttagcga aaattaatag
     5581 aattatattc gcaagacaat tgtgacatta aattaaaaaa ttgtagaatt tttactatcc
     5641 tctttaacgc catgagcctt tataaaaagg ttaaattagt tttaacattc tttttttgag
     5701 taagagttta caatttatca aaacctgtgt tattatattc attagtttca atttattagc
     5761 atctagagaa aaatcaattg gcagttacat tgttggaatt tatgaagaaa aagaatctac
     5821 aacggagaat aagttgctga tcgctttccc taaaattgta tattttgctg agcttatttt
     5881 gacatttcgt tgaagttttc tctaattcgc attcatttta agtaaaacaa tgagaaataa
     5941 aattacaaaa aatacaaatt aaaatacaat ttttagctat aattatagac gatgcccttg
     6001 tatcccattc tgtctcgctt gcccctactt tttatctttt atataccata atgaacgctg
     6061 ccgctactaa ccataccccg attttacatt tcggactccc aaggacgtac aaaatagaaa
     6121 actatagaaa aaaataatca gaaaatagca tgtcatctct ttgtaaaacg cgtttgcaag
     6181 aagaaaggaa acaatggaga agagatcatc catttgtatg taaattttag taaacttgaa
     6241 gaaatcacta acaacttctc ttacttaggg attctatgca aaaccttgta aatcatctga
     6301 tggaggactc gatttaatga attggaaggt tggatttcca ggtggcactt ttcggggaaa
     6361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat
     7201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7621 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7861 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
