LOCUS       sequence-180408-        4223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe General Purpose Vector - with empty BsaI-pad for new potential
            markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4223)
  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 4223)
  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: pYZ189
FEATURES             Location/Qualifiers
     source          1..4223
                     /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..795
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISRLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     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      578..1033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(665..684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     875..896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      1159..1942
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        2318..2422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2423..2491,2492..3283)
                     /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(2641..2660)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3454..4042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3943..3962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4196..4213
                     /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 tggcgcgacg cgccctgtag cggcgcatta
      601 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      661 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      721 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      781 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      841 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
      901 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
      961 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1021 acgtttacaa tttagagacc ggttctttgg gtcaagaccg ggtctctgtc gatcgactct
     1081 agaggaaatt cgagtctaac tccttaacca ctcggacata gtgacttatc tgacactcat
     1141 tgtaaattaa tatatatata ggcattttgt ttagttaaag gtacttaagt aattagtata
     1201 aacgaaccaa ttttataatc aggaagttaa gtgaatggta gcacatgtcg taaaaattgt
     1261 gaatttttat tgaataatat tttaaataca agcctttcta gactaggtat actcataaac
     1321 atatatgagc aaaaggatag aggagattac attgcatctt ctacaaattt atttattgcc
     1381 ctttactgaa aaattaaata atgagtacta aatgataaaa agcgctcagt acaagaaaga
     1441 ttgcaaaaat attgcattct tcatgaatta aagttgcata taaggcatat tgaaagtaat
     1501 agtactaaaa cagcagttag cgaaaattaa tagaattata ttcgcaagac aattgtgaca
     1561 ttaaattaaa aaattgtaga atttttacta tcctctttaa cgccatgagc ctttataaaa
     1621 aggttaaatt agttttaaca ttcttttttt gagtaagagt ttacaattta tcaaaacctg
     1681 tgttattata ttcattagtt tcaatttatt agcatctaga gaaaaatcaa ttggcagtta
     1741 cattgttgga atttatgaag aaaaagaatc tacaacggag aataagttgc tgatcgcttt
     1801 ccctaaaatt gtatattttg ctgagcttat tttgacattt cgttgaagtt ttctctaatt
     1861 cgcattcatt ttaagtaaaa caatgagaaa taaaattaca aaaaatacaa attaaaatac
     1921 aatttttagc tataattata gacgatgccc ttgtatccca ttctgtctcg cttgccccta
     1981 ctttttatct tttatatacc ataatgaacg ctgccgctac taaccatacc ccgattttac
     2041 atttcggact cccaaggacg tacaaaatag aaaactatag aaaaaaataa tcagaaaata
     2101 gcatgtcatc tctttgtaaa acgcgtttgc aagaagaaag gaaacaatgg agaagagatc
     2161 atccatttgt atgtaaattt tagtaaactt gaagaaatca ctaacaactt ctcttactta
     2221 gggattctat gcaaaacctt gtaaatcatc tgatggagga ctcgatttaa tgaattggaa
     2281 ggttggattt ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2341 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2401 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2461 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2521 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2581 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2641 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2701 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2761 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2821 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2881 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2941 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3001 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3061 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3121 tggagccggt gagcgtggat ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3181 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3241 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3301 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3361 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3421 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3481 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3541 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3601 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3661 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3721 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3781 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3841 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3901 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3961 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4021 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4081 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4141 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4201 gtcagtgagc gaggaagcgg aag
//
