LOCUS       sequence-199359-        6406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt81 promoter with empty BsaI-pad - no 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  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 6406)
  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: pYZ184
FEATURES             Location/Qualifiers
     source          1..6406
                     /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..2244
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     rep_origin      3342..4125
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4501..4605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4606..4674,4675..5466)
                     /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(4824..4843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5637..6225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6126..6145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6379..6396
                     /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 ttgaagatgg aagaggaatc
     2161 ctggcatatc atcaattgaa taagttgaat taattatttc aatctcattc tcactttctg
     2221 acttatagtc gctttgttaa atcatatgtc gactctagag gatccccggg taaaaggaat
     2281 gtctcccttg ccagtactgc tagggttttt ctttcaaact atggaagccc attcaagctg
     2341 catattacga ttttgttttt cgcttttaga aagtggttta gatgagataa tagaaaaatt
     2401 cttgatctcc gacaacgagt acttttattt tttttgctaa tcactttact caatattagc
     2461 tcgaaatcgt agaaacgtag acgggtgcgg gataccgagt ggtgtagtta agaattttta
     2521 taaaccacgt ggcccaaaaa tatgaaccca aaacgtttat acatgagtat actttaagaa
     2581 ggctataccc cttcgtgtta gatgtagttt tagctaccca acccgagtct atgagcttga
     2641 cttcagatgt agaaggcatt aaatcgtttt gaatattaat taaaaaacga tgaaaattaa
     2701 atatttaaaa gcaatcatac gctgaaaatt tagtgctgtg gctaatcctt caacatggaa
     2761 atgccataaa agtgactttg acaaaaaaaa aagtatatac aggtagtaaa ctcatctact
     2821 tcattgactt tgtttacagc atgtggaagg aggaatattt attgctaaat cgtagtttaa
     2881 cattcaataa gtaatactat tgaaattcga caagattggc cgcatggatg aaaaagaggc
     2941 attttgcttt gggagaatta gtccaaatta gaactgaaaa aaaaaacttt acgaggcaaa
     3001 aatgtcggat tgagatcgta aaagttcgct cgtcgtcttt tgctttgtga ttgttttcat
     3061 ggatacatct tgctggatat ttaaatttta gtactatgta taagatattc tataaatgtt
     3121 ttatcaccca aacctgttag cgccttctta attctattca atctggcttt tgctctgaga
     3181 ctacttcttg gactttcact acttgttagt tatacggaat ttgtgtaatt agaagtgaaa
     3241 taatcctttc tattagtaat gcgagctgca attcgagtct aactccttaa ccactcggac
     3301 atagtgactt atctgacact cattgtaaat taatatatat ataggcattt tgtttagtta
     3361 aaggtactta agtaattagt ataaacgaac caattttata atcaggaagt taagtgaatg
     3421 gtagcacatg tcgtaaaaat tgtgaatttt tattgaataa tattttaaat acaagccttt
     3481 ctagactagg tatactcata aacatatatg agcaaaagga tagaggagat tacattgcat
     3541 cttctacaaa tttatttatt gccctttact gaaaaattaa ataatgagta ctaaatgata
     3601 aaaagcgctc agtacaagaa agattgcaaa aatattgcat tcttcatgaa ttaaagttgc
     3661 atataaggca tattgaaagt aatagtacta aaacagcagt tagcgaaaat taatagaatt
     3721 atattcgcaa gacaattgtg acattaaatt aaaaaattgt agaattttta ctatcctctt
     3781 taacgccatg agcctttata aaaaggttaa attagtttta acattctttt tttgagtaag
     3841 agtttacaat ttatcaaaac ctgtgttatt atattcatta gtttcaattt attagcatct
     3901 agagaaaaat caattggcag ttacattgtt ggaatttatg aagaaaaaga atctacaacg
     3961 gagaataagt tgctgatcgc tttccctaaa attgtatatt ttgctgagct tattttgaca
     4021 tttcgttgaa gttttctcta attcgcattc attttaagta aaacaatgag aaataaaatt
     4081 acaaaaaata caaattaaaa tacaattttt agctataatt atagacgatg cccttgtatc
     4141 ccattctgtc tcgcttgccc ctacttttta tcttttatat accataatga acgctgccgc
     4201 tactaaccat accccgattt tacatttcgg actcccaagg acgtacaaaa tagaaaacta
     4261 tagaaaaaaa taatcagaaa atagcatgtc atctctttgt aaaacgcgtt tgcaagaaga
     4321 aaggaaacaa tggagaagag atcatccatt tgtatgtaaa ttttagtaaa cttgaagaaa
     4381 tcactaacaa cttctcttac ttagggattc tatgcaaaac cttgtaaatc atctgatgga
     4441 ggactcgatt taatgaattg gaaggttgga tttccaggtg gcacttttcg gggaaatgtg
     4501 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4561 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4621 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4681 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4741 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4801 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4861 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4921 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4981 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5041 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5101 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5161 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5221 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5281 ggctggttta ttgctgataa atctggagcc ggtgagcgtg gatctcgcgg tatcattgca
     5341 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5401 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5461 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5521 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5581 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5761 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6301 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6361 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
