LOCUS       sequence-199358-        6408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt41 promoter with empty BsaI-pad - no 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6408)
  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 6408)
  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: pYZ183
FEATURES             Location/Qualifiers
     source          1..6408
                     /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        1093..2247
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      3344..4127
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4503..4607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4608..4676,4677..5468)
                     /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(4826..4845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5639..6227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6128..6147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6381..6398
                     /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 ttacaattta gagaccggtt ctttggtcaa gaccgggtct ctgtcgatcg actctagagg
     1081 atcagaaaat tatcgccata aaagacagaa taagtcatca gcggttgttt catttcctat
     1141 attttttttt tattttttta ttttttaata agggaaaatt taacgtctaa ggatacagaa
     1201 gattgttagc acattaaagt aataaaggct taagtagtaa gtgccttagc atgttattgt
     1261 atttcaaagg acataatcta aaataataac aatatcattt ctcacaagtt attcaatttt
     1321 cttttttttt tctaataata tcaagaatgt attatttgtt tgacataagt caactaattt
     1381 atttaatatg ctggattaat cttgcagaca tgtaaattaa caagttttag tcaaataacg
     1441 ttgaagtttc aatgaactca aataatttct cttttttttt atataaccat aatctgattt
     1501 atattttccg cagggatcaa ctgaagttat gacatttgga ttggatcact tataaccttg
     1561 gtcgccaaat aatacaaaaa tcagcgttat aaaacaaaga aggtttttgt taagaaatta
     1621 atcctctttc ttgataagaa agttgaaccg aaattgcaga tactgatata tgaaaataat
     1681 acccacaatt ttgggaatag cgcaagcctc aatttaaaca ataggtgagg acacatgata
     1741 atgacctcaa tgattgttag aagaaaagag cctcattaca aaatcgaaaa atgaatggtt
     1801 gggtacaagt ttccaaaaca tggtaaagtg gactttgcgt atgagacgta aatagaaaaa
     1861 aacacttgtt atatgttttc tagaattatt gttgtctctt tatggttgga tgatgcaaaa
     1921 tagtaatttc ggttagttgc tgtaaaacac cacgagacaa atagatatgg atatttatta
     1981 aatcaggaaa aacgtaactc tcggctactg gatggttcag tcacccaacg attactgggg
     2041 agagaaaaca gggcaaaagc aaagcttaaa ggaatccgat tgtcattcgg caatgtgcag
     2101 cgaaactaaa aaccggataa tggacctgtt aatcgaaaca ttgaagataa aggaagagga
     2161 atcctggcat atcatcaatt gaataagttg aattaattat ttcaatctca ttctcacttt
     2221 ctgacttata gtcgctttgt taaatcatat gtcgactcta gaggatcccc gggtaaaagg
     2281 aatgtctccc ttgccagtac tgctagggtt tttctttcaa actatggaag cccattcaag
     2341 ctgcatatta cgattttgtt tttcgctttt agaaagtggt ttagatgaga taatagaaaa
     2401 attcttgatc tccgacaacg agtactttta ttttttttgc taatcacttt actcaatatt
     2461 agctcgaaat cgtagaaacg tagacgggtg cgggataccg agtggtgtag ttaagaattt
     2521 ttataaacca cgtggcccaa aaatatgaac ccaaaacgtt tatacatgag tatactttaa
     2581 gaaggctata ccccttcgtg ttagatgtag ttttagctac ccaacccgag tctatgagct
     2641 tgacttcaga tgtagaaggc attaaatcgt tttgaatatt aattaaaaaa cgatgaaaat
     2701 taaatattta aaagcaatca tacgctgaaa atttagtgct gtggctaatc cttcaacatg
     2761 gaaatgccat aaaagtgact ttgacaaaaa aaaaagtata tacaggtagt aaactcatct
     2821 acttcattga ctttgtttac agcatgtgga aggaggaata tttattgcta aatcgtagtt
     2881 taacattcaa taagtaatac tattgaaatt cgacaagatt ggccgcatgg atgaaaaaga
     2941 ggcattttgc tttgggagaa ttagtccaaa ttagaactga aaaaaaaact ttacgaggca
     3001 aaaatgtcgg attgagatcg taaaagttcg ctcgtcgtct tttgctttgt gattgttttc
     3061 atggatacat cttgctggat atttaaattt tagtactatg tataagatat tctataaatg
     3121 ttttatcacc caaacctgtt agcgccttct taattctatt caatctggct tttgctctga
     3181 gactacttct tggactttca ctacttgtta gttatacgga atttgtgtaa ttagaagtga
     3241 aataatcctt tctattagta atgcgagctg caattcgagt ctaactcctt aaccactcgg
     3301 acatagtgac ttatctgaca ctcattgtaa attaatatat atataggcat tttgtttagt
     3361 taaaggtact taagtaatta gtataaacga accaatttta taatcaggaa gttaagtgaa
     3421 tggtagcaca tgtcgtaaaa attgtgaatt tttattgaat aatattttaa atacaagcct
     3481 ttctagacta ggtatactca taaacatata tgagcaaaag gatagaggag attacattgc
     3541 atcttctaca aatttattta ttgcccttta ctgaaaaatt aaataatgag tactaaatga
     3601 taaaaagcgc tcagtacaag aaagattgca aaaatattgc attcttcatg aattaaagtt
     3661 gcatataagg catattgaaa gtaatagtac taaaacagca gttagcgaaa attaatagaa
     3721 ttatattcgc aagacaattg tgacattaaa ttaaaaaatt gtagaatttt tactatcctc
     3781 tttaacgcca tgagccttta taaaaaggtt aaattagttt taacattctt tttttgagta
     3841 agagtttaca atttatcaaa acctgtgtta ttatattcat tagtttcaat ttattagcat
     3901 ctagagaaaa atcaattggc agttacattg ttggaattta tgaagaaaaa gaatctacaa
     3961 cggagaataa gttgctgatc gctttcccta aaattgtata ttttgctgag cttattttga
     4021 catttcgttg aagttttctc taattcgcat tcattttaag taaaacaatg agaaataaaa
     4081 ttacaaaaaa tacaaattaa aatacaattt ttagctataa ttatagacga tgcccttgta
     4141 tcccattctg tctcgcttgc ccctactttt tatcttttat ataccataat gaacgctgcc
     4201 gctactaacc ataccccgat tttacatttc ggactcccaa ggacgtacaa aatagaaaac
     4261 tatagaaaaa aataatcaga aaatagcatg tcatctcttt gtaaaacgcg tttgcaagaa
     4321 gaaaggaaac aatggagaag agatcatcca tttgtatgta aattttagta aacttgaaga
     4381 aatcactaac aacttctctt acttagggat tctatgcaaa accttgtaaa tcatctgatg
     4441 gaggactcga tttaatgaat tggaaggttg gatttccagg tggcactttt cggggaaatg
     4501 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4561 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4621 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4681 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4741 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4801 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4861 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4921 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4981 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5041 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5101 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5161 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5221 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5281 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggatctcgc ggtatcattg
     5341 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5401 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5461 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5521 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5581 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5641 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5701 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
