LOCUS       sequence-199439-        6368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe General Purpose Vector - with Sp.lys9 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6368)
  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 6368)
  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: pYZ193
FEATURES             Location/Qualifiers
     source          1..6368
                     /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"
     RBS             3112..3123
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      3304..4087
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4463..4567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4568..4636,4637..5428)
                     /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(4786..4805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5599..6187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6088..6107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6341..6358
                     /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 ttacaatttt gttcaaataa tcgagaagag gttatttttt cttcgagttt tacgtcggtt
     1081 acctaagact aaatacatct acttagaaaa tacgaaaacc ccaagtaaag cttcacataa
     1141 ataaggttgc aaaacaaact gaacatttat acttatgaac atgtagataa attgccaaaa
     1201 aaaaggttga cgaactctaa gattacgaat aattatagaa aatgaagaag ctttggtgtt
     1261 ctaaaatgtc aaacttatat ataagagttt ggaaccaaag cgtgaaacat gcttactctt
     1321 cgtcaataat ttcttcctcc aatcgaatac cttctttggc aagtgtatca atcaatggac
     1381 cgcaaagctt catatcatta ggagccaaaa cacctggggt gttaataaca ccatcaagaa
     1441 tttgctgagt agcaacacca caaggaacgc cgactaattt tgccatagag gtgtagccat
     1501 taggaacacc gtagtcaagc aaagtgcaag tacgagtttg gcgttttcct tctttggttt
     1561 caacttcaaa cttgtgttga aggattaaca tatcacgttc accttcttcg tattgcataa
     1621 gttcctccaa ggtagcgcat aaggtgtcca aaggattacc acgaggagta acgggtttgc
     1681 tagagaacat accaagccac ttcaaaccat tcaaaatgcg cttcttgtcg tcttcatctt
     1741 tgaatttgtg gatggaagaa attttcttga ttaggtcggc ttcggaggaa gattcagcct
     1801 tgatgacacg agcagtaacc tccttccaag gcaaagcggg tgcttcagga gataagtatt
     1861 cctgagctgt ctcatccaaa aatcccatat caaccaaaca atgaataaac tcgggaaatc
     1921 cttggtaacg taaagtaccg cgaatgatag tttctgcttc aggtatttga tagcgctctt
     1981 ggtagacagt ggaatcacga ttaggataac aaacgaaagc gtaaccggga tagatgaaat
     2041 agggtttagc agtttccatc aaatccttgc cgtcaatttc aacaagtttg ccgttttcat
     2101 aaaacttggc agagttgcga agagccaaca aaacaccacg agaagaccaa gaaaacttgt
     2161 atcctaaagg attgttggag tcctcgggag cgggaagacc tccacaataa gacaagaaag
     2221 acttaatctt accgccagct ttatgaacct cttcaatggt cttgatagca tataaatgat
     2281 caataccggg atcgacgccg atttcgttca tacaaataga accagccttg atggcagctt
     2341 cctccaactc agccattttg ggattcacat aactggtggt acaaacgtgt ttaccatgct
     2401 taatggcagc tttcataact gttgcatgat aagtgtaagg aatcaagctg atggtaagat
     2461 catgttcgga aacagctttc tcaagagctg cttcatcatt gacatccaaa gcaatggctt
     2521 tgctgtttgg gataccatta ataaatgctt cagccttaga aagagttctg catgctacgg
     2581 ttatattatt ttctttacga cgagataaat attctaaagt aggatgagcg acaaatcccg
     2641 aaccaagtaa taagatggaa ggcattttga tgttgaaaat gaaagaggca gtcagttaag
     2701 gacaagtggt aaaccaaaga cttctgtttt gtcgtcacgg ttttagcgat atataagggg
     2761 ttccttttta tccacagatt tatttctgat ggccgaaatc gtcgtctaaa ttgatgaaaa
     2821 ctgattatct cgaatcggtt aaaaaagcag tcacatggac cattttttga attaatattt
     2881 aatcgttacg tttcctctca atgttatgtt tggcttacta ggggaattgg ttattatcga
     2941 caagtttagt cgataatttg ttagcagcga tgtcgcaact gatagtttta gcaacttttt
     3001 ttacgtagtt tttttctgca caacagaagt tgcttaaaag ctagtagatg ccattcgttg
     3061 ttgtttgggg gtagattaaa gaattgtaag ctttagtata agaataattt gtttctcctc
     3121 tttctgccta tctgttgttc ataatcaact gcgtagttct ggtgaaggag taaattaaac
     3181 tgcctcgcac cgccgtactc atttttacct cggtcgatcg actctagagg aaattcgagt
     3241 ctaactcctt aaccactcgg acatagtgac ttatctgaca ctcattgtaa attaatatat
     3301 atataggcat tttgtttagt taaaggtact taagtaatta gtataaacga accaatttta
     3361 taatcaggaa gttaagtgaa tggtagcaca tgtcgtaaaa attgtgaatt tttattgaat
     3421 aatattttaa atacaagcct ttctagacta ggtatactca taaacatata tgagcaaaag
     3481 gatagaggag attacattgc atcttctaca aatttattta ttgcccttta ctgaaaaatt
     3541 aaataatgag tactaaatga taaaaagcgc tcagtacaag aaagattgca aaaatattgc
     3601 attcttcatg aattaaagtt gcatataagg catattgaaa gtaatagtac taaaacagca
     3661 gttagcgaaa attaatagaa ttatattcgc aagacaattg tgacattaaa ttaaaaaatt
     3721 gtagaatttt tactatcctc tttaacgcca tgagccttta taaaaaggtt aaattagttt
     3781 taacattctt tttttgagta agagtttaca atttatcaaa acctgtgtta ttatattcat
     3841 tagtttcaat ttattagcat ctagagaaaa atcaattggc agttacattg ttggaattta
     3901 tgaagaaaaa gaatctacaa cggagaataa gttgctgatc gctttcccta aaattgtata
     3961 ttttgctgag cttattttga catttcgttg aagttttctc taattcgcat tcattttaag
     4021 taaaacaatg agaaataaaa ttacaaaaaa tacaaattaa aatacaattt ttagctataa
     4081 ttatagacga tgcccttgta tcccattctg tctcgcttgc ccctactttt tatcttttat
     4141 ataccataat gaacgctgcc gctactaacc ataccccgat tttacatttc ggactcccaa
     4201 ggacgtacaa aatagaaaac tatagaaaaa aataatcaga aaatagcatg tcatctcttt
     4261 gtaaaacgcg tttgcaagaa gaaaggaaac aatggagaag agatcatcca tttgtatgta
     4321 aattttagta aacttgaaga aatcactaac aacttctctt acttagggat tctatgcaaa
     4381 accttgtaaa tcatctgatg gaggactcga tttaatgaat tggaaggttg gatttccagg
     4441 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4501 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4561 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4621 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4681 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4741 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4801 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4861 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4921 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4981 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5041 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5101 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5161 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5221 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5281 tggatctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5341 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5401 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5461 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5521 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5581 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5641 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5701 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5761 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5821 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5881 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5941 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6001 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6061 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6121 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6181 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6241 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6301 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6361 agcggaag
//
