LOCUS       sequence-180412-        6372 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 6372)
  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 6372)
  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..6372
                     /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"
     RBS             3116..3127
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      3308..4091
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4467..4571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4572..4640,4641..5432)
                     /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(4790..4809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5603..6191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6092..6111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6345..6362
                     /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 ttttgttcaa ataatcgaga agaggttatt ttttcttcga gttttacgtc
     1081 ggttacctaa gactaaatac atctacttag aaaatacgaa aaccccaagt aaagcttcac
     1141 ataaataagg ttgcaaaaca aactgaacat ttatacttat gaacatgtag ataaattgcc
     1201 aaaaaaaagg ttgacgaact ctaagattac gaataattat agaaaatgaa gaagctttgg
     1261 tgttctaaaa tgtcaaactt atatataaga gtttggaacc aaagcgtgaa acatgcttac
     1321 tcttcgtcaa taatttcttc ctccaatcga ataccttctt tggcaagtgt atcaatcaat
     1381 ggaccgcaaa gcttcatatc attaggagcc aaaacacctg gggtgttaat aacaccatca
     1441 agaatttgct gagtagcaac accacaagga acgccgacta attttgccat agaggtgtag
     1501 ccattaggaa caccgtagtc aagcaaagtg caagtacgag tttggcgttt tccttctttg
     1561 gtttcaactt caaacttgtg ttgaaggatt aacatatcac gttcaccttc ttcgtattgc
     1621 ataagttcct ccaaggtagc gcataaggtg tccaaaggat taccacgagg agtaacgggt
     1681 ttgctagaga acataccaag ccacttcaaa ccattcaaaa tgcgcttctt gtcgtcttca
     1741 tctttgaatt tgtggatgga agaaattttc ttgattaggt cggcttcgga ggaagattca
     1801 gccttgatga cacgagcagt aacctccttc caaggcaaag cgggtgcttc aggagataag
     1861 tattcctgag ctgtctcatc caaaaatccc atatcaacca aacaatgaat aaactcggga
     1921 aatccttggt aacgtaaagt accgcgaatg atagtttctg cttcaggtat ttgatagcgc
     1981 tcttggtaga cagtggaatc acgattagga taacaaacga aagcgtaacc gggatagatg
     2041 aaatagggtt tagcagtttc catcaaatcc ttgccgtcaa tttcaacaag tttgccgttt
     2101 tcataaaact tggcagagtt gcgaagagcc aacaaaacac cacgagaaga ccaagaaaac
     2161 ttgtatccta aaggattgtt ggagtcctcg ggagcgggaa gacctccaca ataagacaag
     2221 aaagacttaa tcttaccgcc agctttatga acctcttcaa tggtcttgat agcatataaa
     2281 tgatcaatac cgggatcgac gccgatttcg ttcatacaaa tagaaccagc cttgatggca
     2341 gcttcctcca actcagccat tttgggattc acataactgg tggtacaaac gtgtttacca
     2401 tgcttaatgg cagctttcat aactgttgca tgataagtgt aaggaatcaa gctgatggta
     2461 agatcatgtt cggaaacagc tttctcaaga gctgcttcat cattgacatc caaagcaatg
     2521 gctttgctgt ttgggatacc attaataaat gcttcagcct tagaaagagt tctgcatgct
     2581 acggttatat tattttcttt acgacgagat aaatattcta aagtaggatg agcgacaaat
     2641 cccgaaccaa gtaataagat ggaaggcatt ttgatgttga aaatgaaaga ggcagtcagt
     2701 taaggacaag tggtaaacca aagacttctg ttttgtcgtc acggttttag cgatatataa
     2761 ggggttcctt tttatccaca gatttatttc tgatggccga aatcgtcgtc taaattgatg
     2821 aaaactgatt atctcgaatc ggttaaaaaa gcagtcacat ggaccatttt ttgaattaat
     2881 atttaatcgt tacgtttcct ctcaatgtta tgtttggctt actaggggaa ttggttatta
     2941 tcgacaagtt tagtcgataa tttgttagca gcgatgtcgc aactgatagt tttagcaact
     3001 ttttttacgt agtttttttc tgcacaacag aagttgctta aaagctagta gatgccattc
     3061 gttgttgttt gggggtagat taaagaattg taagctttag tataagaata atttgtttct
     3121 cctctttctg cctatctgtt gttcataatc aactgcgtag ttctggtgaa ggagtaaatt
     3181 aaactgcctc gcaccgccgt actcattttt acctcggtcg atcgactcta gaggaaattc
     3241 gagtctaact ccttaaccac tcggacatag tgacttatct gacactcatt gtaaattaat
     3301 atatatatag gcattttgtt tagttaaagg tacttaagta attagtataa acgaaccaat
     3361 tttataatca ggaagttaag tgaatggtag cacatgtcgt aaaaattgtg aatttttatt
     3421 gaataatatt ttaaatacaa gcctttctag actaggtata ctcataaaca tatatgagca
     3481 aaaggataga ggagattaca ttgcatcttc tacaaattta tttattgccc tttactgaaa
     3541 aattaaataa tgagtactaa atgataaaaa gcgctcagta caagaaagat tgcaaaaata
     3601 ttgcattctt catgaattaa agttgcatat aaggcatatt gaaagtaata gtactaaaac
     3661 agcagttagc gaaaattaat agaattatat tcgcaagaca attgtgacat taaattaaaa
     3721 aattgtagaa tttttactat cctctttaac gccatgagcc tttataaaaa ggttaaatta
     3781 gttttaacat tctttttttg agtaagagtt tacaatttat caaaacctgt gttattatat
     3841 tcattagttt caatttatta gcatctagag aaaaatcaat tggcagttac attgttggaa
     3901 tttatgaaga aaaagaatct acaacggaga ataagttgct gatcgctttc cctaaaattg
     3961 tatattttgc tgagcttatt ttgacatttc gttgaagttt tctctaattc gcattcattt
     4021 taagtaaaac aatgagaaat aaaattacaa aaaatacaaa ttaaaataca atttttagct
     4081 ataattatag acgatgccct tgtatcccat tctgtctcgc ttgcccctac tttttatctt
     4141 ttatatacca taatgaacgc tgccgctact aaccataccc cgattttaca tttcggactc
     4201 ccaaggacgt acaaaataga aaactataga aaaaaataat cagaaaatag catgtcatct
     4261 ctttgtaaaa cgcgtttgca agaagaaagg aaacaatgga gaagagatca tccatttgta
     4321 tgtaaatttt agtaaacttg aagaaatcac taacaacttc tcttacttag ggattctatg
     4381 caaaaccttg taaatcatct gatggaggac tcgatttaat gaattggaag gttggatttc
     4441 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4561 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4621 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4681 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4741 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4801 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4861 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4921 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4981 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5041 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5101 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5161 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5221 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5281 agcgtggatc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5341 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5401 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5461 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag
     6001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6301 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6361 aggaagcgga ag
//
