LOCUS       sequence-199432-        5943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe General Purpose Vector - with Sp.ura4 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5943)
  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 5943)
  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: pYZ190
FEATURES             Location/Qualifiers
     source          1..5943
                     /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="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     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     297..313
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     297..313
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(347..363)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(348..364)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     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"
     CDS             complement(1465..2259)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     rep_origin      2879..3662
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4038..4142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4143..4211,4212..5003)
                     /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(4361..4380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5174..5762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5663..5682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5916..5933
                     /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 gaactagtgg atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc
      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 gcttgtgata ttgacgaaac tttttgacat ctaatttatt ctgttccaac
     1081 accaatgttt ataaccaagt tttatcttgt ttgtctacat ggtattttac attcatctac
     1141 atacatcttt cattggcttt gtacatagtt atcattacaa gtctaaaaaa attcactctt
     1201 ttcttattca atgtcaatcc aagagaaaag attgtggtaa tgttgtagga gcatgtttaa
     1261 taaattacta tagcaaatta ctttttattc ccaaggtgtt tatctataat agttaatatt
     1321 ttagtcgcta cataaaattt taccaaagag tacttgtata ctaattctaa atgccttctg
     1381 acataaaacg cctaggaaaa caaacgcaaa caaggcatcg actttttcaa taaccaacca
     1441 aaaaaatttt acattagtct ttttttaatg ctgagaaagt ctttgctgat atgccttcca
     1501 accagcttct ctatatctct tggcttcgac aacaggatta cgaccagctc catagactcc
     1561 acgaccaaca atgatgatat cgctaccgca gtttacaatc acttcttcag gagtacgata
     1621 ttgctgtccc agcccgtctc ctttaacatc caagccgata ccaggggaca tagttatgta
     1681 gtcgctttga aggttaggaa atcgacgacc agctataaag ccaaagcaaa aatcggtatg
     1741 cttctcaaac cattctaagg ttttctctgt gtaggaacca gtagccaaag agcctttgga
     1801 agacatttca gccaaaagca agagtccacg tcccaaaggt aaaccaactt ctttgaggcc
     1861 ttgtataata ccctcgcctg gcactgtatg gcaatttgtg atatgagccc aagaagcaat
     1921 tttgtacaca ccagatgcat attgtagctt gacggtattt ccaatgtctg cgaatttgcg
     1981 atcctcaaag ataagaaaac gatgcttttt acctaaggcc accagttttt ctaccatatc
     2041 ctggtcgaaa tcctcgacaa cgtcaatatg tgtcttgata acacagacat agggtccaat
     2101 tttatctacc aattctaaga tttcggattt cttcgtcaaa tcgaccgcga ctgacaagtt
     2161 gctttgcttt tcttccatca aagccaacaa ttccttggca atgggatttt tcatcccctc
     2221 agctctagct gaatagcttt gaaatactct agcatccata actttgcttt taaaccttta
     2281 atttcgatcc aagcaaaaaa gaggttcttg gtaggacaat acggtaagaa aacacgacat
     2341 gtgcagagat gccgacgaag catagttaaa ctgggatggt aaaatcaatt aagaatttat
     2401 aaagacaaaa ttgtataagt ctctaaaaca tcttaattat acctcacaga actatctaaa
     2461 atatattcac aaagtgcaaa cattatcatg aaaaagaacc attttaattt aaagcaaggg
     2521 cattaaggct tatttacaga atttcttact tttgtaaaga ttataaggct gattatcttt
     2581 ttcaccatgc caaaaattac acaagataga atggatgttt gaaattaaac gtgagtatac
     2641 aaacaaatac actaggtaaa tcgaaacatt tttttctcca ttaagtaaca aattcctatt
     2701 tagagaaaga atgctgagta gattaaataa tctatacaaa cttttttaac acaaatgcat
     2761 acatatagcc agtgggattt gtagctagtc gatcgactct agaggaaatt cgagtctaac
     2821 tccttaacca ctcggacata gtgacttatc tgacactcat tgtaaattaa tatatatata
     2881 ggcattttgt ttagttaaag gtacttaagt aattagtata aacgaaccaa ttttataatc
     2941 aggaagttaa gtgaatggta gcacatgtcg taaaaattgt gaatttttat tgaataatat
     3001 tttaaataca agcctttcta gactaggtat actcataaac atatatgagc aaaaggatag
     3061 aggagattac attgcatctt ctacaaattt atttattgcc ctttactgaa aaattaaata
     3121 atgagtacta aatgataaaa agcgctcagt acaagaaaga ttgcaaaaat attgcattct
     3181 tcatgaatta aagttgcata taaggcatat tgaaagtaat agtactaaaa cagcagttag
     3241 cgaaaattaa tagaattata ttcgcaagac aattgtgaca ttaaattaaa aaattgtaga
     3301 atttttacta tcctctttaa cgccatgagc ctttataaaa aggttaaatt agttttaaca
     3361 ttcttttttt gagtaagagt ttacaattta tcaaaacctg tgttattata ttcattagtt
     3421 tcaatttatt agcatctaga gaaaaatcaa ttggcagtta cattgttgga atttatgaag
     3481 aaaaagaatc tacaacggag aataagttgc tgatcgcttt ccctaaaatt gtatattttg
     3541 ctgagcttat tttgacattt cgttgaagtt ttctctaatt cgcattcatt ttaagtaaaa
     3601 caatgagaaa taaaattaca aaaaatacaa attaaaatac aatttttagc tataattata
     3661 gacgatgccc ttgtatccca ttctgtctcg cttgccccta ctttttatct tttatatacc
     3721 ataatgaacg ctgccgctac taaccatacc ccgattttac atttcggact cccaaggacg
     3781 tacaaaatag aaaactatag aaaaaaataa tcagaaaata gcatgtcatc tctttgtaaa
     3841 acgcgtttgc aagaagaaag gaaacaatgg agaagagatc atccatttgt atgtaaattt
     3901 tagtaaactt gaagaaatca ctaacaactt ctcttactta gggattctat gcaaaacctt
     3961 gtaaatcatc tgatggagga ctcgatttaa tgaattggaa ggttggattt ccaggtggca
     4021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4141 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4201 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4261 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4321 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4381 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4441 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4501 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4561 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4621 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4681 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4741 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4801 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat
     4861 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4921 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4981 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5041 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5101 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5161 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5221 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5281 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5341 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5401 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5461 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5521 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5581 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5641 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5701 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5761 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5821 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5881 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5941 aag
//
