LOCUS       sequence-207094-        8268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt81 promoter with Sp.leu1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8268)
  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 8268)
  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: pYZ160
FEATURES             Location/Qualifiers
     source          1..8268
                     /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..4106
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     rep_origin      5204..5987
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6363..6467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6468..6536,6537..7328)
                     /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(6686..6705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7499..8087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7988..8007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8241..8258
                     /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 ggaagaggaa
     4021 tcctggcata tcatcaattg aataagttga attaattatt tcaatctcat tctcactttc
     4081 tgacttatag tcgctttgtt aaatcatatg tcgactctag aggatccccg ggtaaaagga
     4141 atgtctccct tgccagtact gctagggttt ttctttcaaa ctatggaagc ccattcaagc
     4201 tgcatattac gattttgttt ttcgctttta gaaagtggtt tagatgagat aatagaaaaa
     4261 ttcttgatct ccgacaacga gtacttttat tttttttgct aatcacttta ctcaatatta
     4321 gctcgaaatc gtagaaacgt agacgggtgc gggataccga gtggtgtagt taagaatttt
     4381 tataaaccac gtggcccaaa aatatgaacc caaaacgttt atacatgagt atactttaag
     4441 aaggctatac cccttcgtgt tagatgtagt tttagctacc caacccgagt ctatgagctt
     4501 gacttcagat gtagaaggca ttaaatcgtt ttgaatatta attaaaaaac gatgaaaatt
     4561 aaatatttaa aagcaatcat acgctgaaaa tttagtgctg tggctaatcc ttcaacatgg
     4621 aaatgccata aaagtgactt tgacaaaaaa aaaagtatat acaggtagta aactcatcta
     4681 cttcattgac tttgtttaca gcatgtggaa ggaggaatat ttattgctaa atcgtagttt
     4741 aacattcaat aagtaatact attgaaattc gacaagattg gccgcatgga tgaaaaagag
     4801 gcattttgct ttgggagaat tagtccaaat tagaactgaa aaaaaaaact ttacgaggca
     4861 aaaatgtcgg attgagatcg taaaagttcg ctcgtcgtct tttgctttgt gattgttttc
     4921 atggatacat cttgctggat atttaaattt tagtactatg tataagatat tctataaatg
     4981 ttttatcacc caaacctgtt agcgccttct taattctatt caatctggct tttgctctga
     5041 gactacttct tggactttca ctacttgtta gttatacgga atttgtgtaa ttagaagtga
     5101 aataatcctt tctattagta atgcgagctg caattcgagt ctaactcctt aaccactcgg
     5161 acatagtgac ttatctgaca ctcattgtaa attaatatat atataggcat tttgtttagt
     5221 taaaggtact taagtaatta gtataaacga accaatttta taatcaggaa gttaagtgaa
     5281 tggtagcaca tgtcgtaaaa attgtgaatt tttattgaat aatattttaa atacaagcct
     5341 ttctagacta ggtatactca taaacatata tgagcaaaag gatagaggag attacattgc
     5401 atcttctaca aatttattta ttgcccttta ctgaaaaatt aaataatgag tactaaatga
     5461 taaaaagcgc tcagtacaag aaagattgca aaaatattgc attcttcatg aattaaagtt
     5521 gcatataagg catattgaaa gtaatagtac taaaacagca gttagcgaaa attaatagaa
     5581 ttatattcgc aagacaattg tgacattaaa ttaaaaaatt gtagaatttt tactatcctc
     5641 tttaacgcca tgagccttta taaaaaggtt aaattagttt taacattctt tttttgagta
     5701 agagtttaca atttatcaaa acctgtgtta ttatattcat tagtttcaat ttattagcat
     5761 ctagagaaaa atcaattggc agttacattg ttggaattta tgaagaaaaa gaatctacaa
     5821 cggagaataa gttgctgatc gctttcccta aaattgtata ttttgctgag cttattttga
     5881 catttcgttg aagttttctc taattcgcat tcattttaag taaaacaatg agaaataaaa
     5941 ttacaaaaaa tacaaattaa aatacaattt ttagctataa ttatagacga tgcccttgta
     6001 tcccattctg tctcgcttgc ccctactttt tatcttttat ataccataat gaacgctgcc
     6061 gctactaacc ataccccgat tttacatttc ggactcccaa ggacgtacaa aatagaaaac
     6121 tatagaaaaa aataatcaga aaatagcatg tcatctcttt gtaaaacgcg tttgcaagaa
     6181 gaaaggaaac aatggagaag agatcatcca tttgtatgta aattttagta aacttgaaga
     6241 aatcactaac aacttctctt acttagggat tctatgcaaa accttgtaaa tcatctgatg
     6301 gaggactcga tttaatgaat tggaaggttg gatttccagg tggcactttt cggggaaatg
     6361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6481 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6541 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6601 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6661 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6721 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6781 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6841 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6901 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6961 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7021 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7081 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7141 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggatctcgc ggtatcattg
     7201 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7261 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7321 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7381 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7441 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7501 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7561 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7621 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7681 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7741 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7801 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7861 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7921 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7981 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8041 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8101 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8161 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8221 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
