LOCUS       sequence-180409-        5947 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 5947)
  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 5947)
  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..5947
                     /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"
     CDS             complement(1469..2263)
                     /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      2883..3666
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4042..4146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4147..4215,4216..5007)
                     /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(4365..4384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5178..5766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5667..5686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5920..5937
                     /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 tttagcttgt gatattgacg aaactttttg acatctaatt tattctgttc
     1081 caacaccaat gtttataacc aagttttatc ttgtttgtct acatggtatt ttacattcat
     1141 ctacatacat ctttcattgg ctttgtacat agttatcatt acaagtctaa aaaaattcac
     1201 tcttttctta ttcaatgtca atccaagaga aaagattgtg gtaatgttgt aggagcatgt
     1261 ttaataaatt actatagcaa attacttttt attcccaagg tgtttatcta taatagttaa
     1321 tattttagtc gctacataaa attttaccaa agagtacttg tatactaatt ctaaatgcct
     1381 tctgacataa aacgcctagg aaaacaaacg caaacaaggc atcgactttt tcaataacca
     1441 accaaaaaaa ttttacatta gtcttttttt aatgctgaga aagtctttgc tgatatgcct
     1501 tccaaccagc ttctctatat ctcttggctt cgacaacagg attacgacca gctccataga
     1561 ctccacgacc aacaatgatg atatcgctac cgcagtttac aatcacttct tcaggagtac
     1621 gatattgctg tcccagcccg tctcctttaa catccaagcc gataccaggg gacatagtta
     1681 tgtagtcgct ttgaaggtta ggaaatcgac gaccagctat aaagccaaag caaaaatcgg
     1741 tatgcttctc aaaccattct aaggttttct ctgtgtagga accagtagcc aaagagcctt
     1801 tggaagacat ttcagccaaa agcaagagtc cacgtcccaa aggtaaacca acttctttga
     1861 ggccttgtat aataccctcg cctggcactg tatggcaatt tgtgatatga gcccaagaag
     1921 caattttgta cacaccagat gcatattgta gcttgacggt atttccaatg tctgcgaatt
     1981 tgcgatcctc aaagataaga aaacgatgct ttttacctaa ggccaccagt ttttctacca
     2041 tatcctggtc gaaatcctcg acaacgtcaa tatgtgtctt gataacacag acatagggtc
     2101 caattttatc taccaattct aagatttcgg atttcttcgt caaatcgacc gcgactgaca
     2161 agttgctttg cttttcttcc atcaaagcca acaattcctt ggcaatggga tttttcatcc
     2221 cctcagctct agctgaatag ctttgaaata ctctagcatc cataactttg cttttaaacc
     2281 tttaatttcg atccaagcaa aaaagaggtt cttggtagga caatacggta agaaaacacg
     2341 acatgtgcag agatgccgac gaagcatagt taaactggga tggtaaaatc aattaagaat
     2401 ttataaagac aaaattgtat aagtctctaa aacatcttaa ttatacctca cagaactatc
     2461 taaaatatat tcacaaagtg caaacattat catgaaaaag aaccatttta atttaaagca
     2521 agggcattaa ggcttattta cagaatttct tacttttgta aagattataa ggctgattat
     2581 ctttttcacc atgccaaaaa ttacacaaga tagaatggat gtttgaaatt aaacgtgagt
     2641 atacaaacaa atacactagg taaatcgaaa catttttttc tccattaagt aacaaattcc
     2701 tatttagaga aagaatgctg agtagattaa ataatctata caaacttttt taacacaaat
     2761 gcatacatat agccagtggg atttgtagct agtcgatcga ctctagagga aattcgagtc
     2821 taactcctta accactcgga catagtgact tatctgacac tcattgtaaa ttaatatata
     2881 tataggcatt ttgtttagtt aaaggtactt aagtaattag tataaacgaa ccaattttat
     2941 aatcaggaag ttaagtgaat ggtagcacat gtcgtaaaaa ttgtgaattt ttattgaata
     3001 atattttaaa tacaagcctt tctagactag gtatactcat aaacatatat gagcaaaagg
     3061 atagaggaga ttacattgca tcttctacaa atttatttat tgccctttac tgaaaaatta
     3121 aataatgagt actaaatgat aaaaagcgct cagtacaaga aagattgcaa aaatattgca
     3181 ttcttcatga attaaagttg catataaggc atattgaaag taatagtact aaaacagcag
     3241 ttagcgaaaa ttaatagaat tatattcgca agacaattgt gacattaaat taaaaaattg
     3301 tagaattttt actatcctct ttaacgccat gagcctttat aaaaaggtta aattagtttt
     3361 aacattcttt ttttgagtaa gagtttacaa tttatcaaaa cctgtgttat tatattcatt
     3421 agtttcaatt tattagcatc tagagaaaaa tcaattggca gttacattgt tggaatttat
     3481 gaagaaaaag aatctacaac ggagaataag ttgctgatcg ctttccctaa aattgtatat
     3541 tttgctgagc ttattttgac atttcgttga agttttctct aattcgcatt cattttaagt
     3601 aaaacaatga gaaataaaat tacaaaaaat acaaattaaa atacaatttt tagctataat
     3661 tatagacgat gcccttgtat cccattctgt ctcgcttgcc cctacttttt atcttttata
     3721 taccataatg aacgctgccg ctactaacca taccccgatt ttacatttcg gactcccaag
     3781 gacgtacaaa atagaaaact atagaaaaaa ataatcagaa aatagcatgt catctctttg
     3841 taaaacgcgt ttgcaagaag aaaggaaaca atggagaaga gatcatccat ttgtatgtaa
     3901 attttagtaa acttgaagaa atcactaaca acttctctta cttagggatt ctatgcaaaa
     3961 ccttgtaaat catctgatgg aggactcgat ttaatgaatt ggaaggttgg atttccaggt
     4021 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4081 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4141 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4201 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4261 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4321 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4381 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4441 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4501 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4561 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4621 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4681 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4741 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4801 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4861 ggatctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4921 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4981 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5041 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5101 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5161 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5221 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5281 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5341 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5401 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5461 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5521 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     5581 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5641 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5701 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5761 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5821 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5881 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5941 gcggaag
//
