LOCUS       sequence-180410-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe General Purpose Vector - with Sp.leu1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  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 6084)
  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: pYZ191
FEATURES             Location/Qualifiers
     source          1..6084
                     /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      3020..3803
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4179..4283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4284..4352,4353..5144)
                     /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(4502..4521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5315..5903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5804..5823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6057..6074
                     /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 tttcaactcg gtcataaagt tgaacggatg tcgtaaatca attccatgct
     1081 tttgcaactt aatacatctt tgcttgacta taacatacca aaagaaaaaa cattagagcc
     1141 ttaaaattta cgcttttgtc aattgcattt ttaacgataa taaatatgcg acacaattaa
     1201 tcaataaaaa ttaggtcaaa tcaaatgata tgaatttatt ctacattaaa ccctattaca
     1261 aaattttttc aagttcttca acaacagcct tagtaatatc agcggtagaa gcctcacctc
     1321 ccaaatcgcg agtataaaga ccacgtccac caattgaagt atcatccaag accttgcgta
     1381 cggcggcttc gatagcttca gcctccttag gagcattcaa accataacga aggagaaggg
     1441 aagcagataa aattgtacca acaggattaa caatgccctt gccagcgata tcgggagcgc
     1501 taccgtgaat gggctcaacc aaacaatgaa ccttttcttc tgattttcct accacaccgg
     1561 aaagggaggc agaaggcaaa aggcccaagc taccaggaat gacagaagcc tcatctgaaa
     1621 taatgtcacc aaacaagttg tcagtcaaaa caacaccgtt aagtgtacga gggctcttga
     1681 ccaaaagcat ggctgcggag tcaatgagct ggttttttaa ggtaaggtga ggatattcct
     1741 ccttaaaaat cttagctaca gtcttgcgcc aaagacgaga agttgccaaa acattagctt
     1801 tgtcgagtaa tgtgacggga gcaggagggt tggaagtttc agctaaccaa gcagccaaac
     1861 gagcaatacg agaaacttct tccaaactgt aaggccaagt gtccatagca taacccgatc
     1921 cgttgtcctc agtgcgctca ccaaagtaac aacctccagt aagttctcgt acaacacaaa
     1981 aatcgacacc ttcaacgatt tcaggcttca aagggctgta cttgactaaa gacttgctgg
     2041 caaagttgca aggtcgaagg ttggcccaaa cacccatact cttacgaagc ttcaataaac
     2101 cttgctcagg acgacaattg gggttggtcc attcaggacc accaacggca cccaaaagaa
     2161 caccgtcagc ttccaaacaa gccttcacag tctcgtcagt caaaggggtt ccataggcat
     2221 caatagaggc acctccaatc ttgtgttctt caaactccag ttttaactca ggtcgcttct
     2281 tctcaacgac tttcaaaacc tccaaggcag aagcaacaat ttcagggcca atatgatctc
     2341 ctggtaagac gacgattttc tttgcacaca tgttgttgaa gaagttttgt tgtgaaatgg
     2401 tttcgtgaaa gtttcagacc ctaccgcaaa aatgcctggt ttcgggaaac tcaacactgt
     2461 tgcacttttt atactacaga ttgggatatc gataatattg cgtaaaaaat ccttttttta
     2521 aaaagcttgt ttacagtaac gtaaatgacc agaaatcaga tgaaaatcac aagaaagcaa
     2581 ataattcacg ttaaatcctg atatgtttga ttttgtgatg aaatcatgga tgttcatagg
     2641 aattgttgaa attgcgcttt tttaacgaaa tatacaagta tcctggagct tacttaatta
     2701 attaatgaat ctttgtttct agatattaaa atagtagcct caattatcag cgctttctac
     2761 gttagtaaac gaaattttta atgtcaaaaa aatgtttaat agacagtacg aatatgcatt
     2821 ataatgttca taaataattt gagtatgtgt acgagttgtc tttaaaccca cagaggtaga
     2881 atgtatatat aaaattaata agctaagtgt aatacttaaa aaatacatgt cgatcgactc
     2941 tagaggaaat tcgagtctaa ctccttaacc actcggacat agtgacttat ctgacactca
     3001 ttgtaaatta atatatatat aggcattttg tttagttaaa ggtacttaag taattagtat
     3061 aaacgaacca attttataat caggaagtta agtgaatggt agcacatgtc gtaaaaattg
     3121 tgaattttta ttgaataata ttttaaatac aagcctttct agactaggta tactcataaa
     3181 catatatgag caaaaggata gaggagatta cattgcatct tctacaaatt tatttattgc
     3241 cctttactga aaaattaaat aatgagtact aaatgataaa aagcgctcag tacaagaaag
     3301 attgcaaaaa tattgcattc ttcatgaatt aaagttgcat ataaggcata ttgaaagtaa
     3361 tagtactaaa acagcagtta gcgaaaatta atagaattat attcgcaaga caattgtgac
     3421 attaaattaa aaaattgtag aatttttact atcctcttta acgccatgag cctttataaa
     3481 aaggttaaat tagttttaac attctttttt tgagtaagag tttacaattt atcaaaacct
     3541 gtgttattat attcattagt ttcaatttat tagcatctag agaaaaatca attggcagtt
     3601 acattgttgg aatttatgaa gaaaaagaat ctacaacgga gaataagttg ctgatcgctt
     3661 tccctaaaat tgtatatttt gctgagctta ttttgacatt tcgttgaagt tttctctaat
     3721 tcgcattcat tttaagtaaa acaatgagaa ataaaattac aaaaaataca aattaaaata
     3781 caatttttag ctataattat agacgatgcc cttgtatccc attctgtctc gcttgcccct
     3841 actttttatc ttttatatac cataatgaac gctgccgcta ctaaccatac cccgatttta
     3901 catttcggac tcccaaggac gtacaaaata gaaaactata gaaaaaaata atcagaaaat
     3961 agcatgtcat ctctttgtaa aacgcgtttg caagaagaaa ggaaacaatg gagaagagat
     4021 catccatttg tatgtaaatt ttagtaaact tgaagaaatc actaacaact tctcttactt
     4081 agggattcta tgcaaaacct tgtaaatcat ctgatggagg actcgattta atgaattgga
     4141 aggttggatt tccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4201 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4261 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4321 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4381 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4441 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4501 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4561 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4621 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4681 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4741 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4801 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4861 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4921 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4981 ctggagccgg tgagcgtgga tctcgcggta tcattgcagc actggggcca gatggtaagc
     5041 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5101 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5161 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5221 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5281 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5341 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5401 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5461 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5521 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5581 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5641 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5701 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5761 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5821 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5881 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5941 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6001 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6061 agtcagtgag cgaggaagcg gaag
//
