LOCUS       Exported                8148 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Pombe Expression Vector - nmt1 promoter with Sp.ura4 marker.
ACCESSION   .
VERSION     .
KEYWORDS    pYZ155
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8148)
  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 8148)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8148
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        143..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"
     promoter        2822..3986
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     rep_origin      5084..5867
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6243..6347
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6348..7208
                     /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(6566..6585)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7379..7967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7868..7887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8121..8138
                     /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 tcagaaaatt
     2821 atcgccataa aagacagaat aagtcatcag cggttgtttc atttcctata tttttttttt
     2881 atttttttat tttttaataa gggaaaattt aacgtctaag gatacagaag attgttagca
     2941 cattaaagta ataaaggctt aagtagtaag tgccttagca tgttattgta tttcaaagga
     3001 cataatctaa aataataaca atatcatttc tcacaagtta ttcaattttc tttttttttt
     3061 ctaataatat caagaatgta ttatttgttt gacataagtc aactaattta tttaatatgc
     3121 tggattaatc ttgcagacat gtaaattaac aagttttagt caaataacgt tgaagtttca
     3181 atgaactcaa ataatttctc ttttttttta tataaccata agtctaatct gatttatatt
     3241 ttccgcaggg atcaactgaa gttatgacat ttggattgga tcacttataa ccttggtcgc
     3301 caaataatac aaaaatcagc gttataaaac aaagaaggtt tttgttaaga aattaatcct
     3361 ctttcttgat aagaaagttg aaccgaaatt gcagatactg atatatgaaa ataataccca
     3421 caattttggg aatagcgcaa gcctcaattt aaacaatagg tgaggacaca tgataatgac
     3481 ctcaatgatt gttagaagaa aagagcctca ttacaaaatc gaaaaatgaa tggttgggta
     3541 caagtttcca aaacatggta aagtggactt tgcgtatgag acgtaaatag aaaaaaacac
     3601 ttgttatatg ttttctagaa ttattgttgt ctctttatgg ttggatgatg caaaatagta
     3661 atttcggtta gttgctgtaa aacaccacga gacaaataga tatggatatt tattaaatca
     3721 ggaaaaacgt aactctcggc tactggatgg ttcagtcacc caacgattac tggggagaga
     3781 aaacagggca aaagcaaagc ttaaaggaat ccgattgtca ttcggcaatg tgcagcgaaa
     3841 ctaaaaaccg gataatggac ctgttaatcg aaacattgaa gatatataaa ggaagaggaa
     3901 tcctggcata tcatcaattg aataagttga attaattatt tcaatctcat tctcactttc
     3961 tgacttatag tcgctttgtt aaatcatatg tcgactctag aggatccccg ggtaaaagga
     4021 atgtctccct tgccagtact gctagggttt ttctttcaaa ctatggaagc ccattcaagc
     4081 tgcatattac gattttgttt ttcgctttta gaaagtggtt tagatgagat aatagaaaaa
     4141 ttcttgatct ccgacaacga gtacttttat tttttttgct aatcacttta ctcaatatta
     4201 gctcgaaatc gtagaaacgt agacgggtgc gggataccga gtggtgtagt taagaatttt
     4261 tataaaccac gtggcccaaa aatatgaacc caaaacgttt atacatgagt atactttaag
     4321 aaggctatac cccttcgtgt tagatgtagt tttagctacc caacccgagt ctatgagctt
     4381 gacttcagat gtagaaggca ttaaatcgtt ttgaatatta attaaaaaac gatgaaaatt
     4441 aaatatttaa aagcaatcat acgctgaaaa tttagtgctg tggctaatcc ttcaacatgg
     4501 aaatgccata aaagtgactt tgacaaaaaa aaaagtatat acaggtagta aactcatcta
     4561 cttcattgac tttgtttaca gcatgtggaa ggaggaatat ttattgctaa atcgtagttt
     4621 aacattcaat aagtaatact attgaaattc gacaagattg gccgcatgga tgaaaaagag
     4681 gcattttgct ttgggagaat tagttcaaat tagaactgaa aaaaaaaact ttacgaggca
     4741 aaaatgtcgg attgagatcg taaaagttcg ctcgtcgtct tttgctttgt gattgttttc
     4801 atggatacat cttgctggat atttaaattt tagtactatg tataagatat tctataaatg
     4861 ttttatcacc caaacctgtt agcgccttct taattctatt caatctggct tttgctctga
     4921 gactacttct tggactttca ctacttgtta gttatacgga atttgtgtaa ttagaagtga
     4981 aataatcctt tctattagta atgcgagctg caattcgagt ctaactcctt aaccactcgg
     5041 acatagtgac ttatctgaca ctcattgtaa attaatatat atataggcat tttgtttagt
     5101 taaaggtact taagtaatta gtataaacga accaatttta taatcaggaa gttaagtgaa
     5161 tggtagcaca tgtcgtaaaa attgtgaatt tttattgaat aatattttaa atacaagcct
     5221 ttctagacta ggtatactca taaacatata tgagcaaaag gatagaggag attacattgc
     5281 atcttctaca aatttattta ttgcccttta ctgaaaaatt aaataatgag tactaaatga
     5341 taaaaagcgc tcagtacaag aaagattgca aaaatattgc attcttcatg aattaaagtt
     5401 gcatataagg catattgaaa gtaatagtac taaaacagca gttagcgaaa attaatagaa
     5461 ttatattcgc aagacaattg tgacattaaa ttaaaaaatt gtagaatttt tactatcctc
     5521 tttaacgcca tgagccttta taaaaaggtt aaattagttt taacattctt tttttgagta
     5581 agagtttaca atttatcaaa acctgtgtta ttatattcat tagtttcaat ttattagcat
     5641 ctagagaaaa atcaattggc agttacattg ttggaattta tgaagaaaaa gaatctacaa
     5701 cggagaataa gttgctgatc gctttcccta aaattgtata ttttgctgag cttattttga
     5761 catttcgttg aagttttctc taattcgcat tcattttaag taaaacaatg agaaataaaa
     5821 ttacaaaaaa tacaaattaa aatacaattt ttagctataa ttatagacga tgcccttgta
     5881 tcccattctg tctcgcttgc ccctactttt tatcttttat ataccataat gaacgctgcc
     5941 gctactaacc ataccccgat tttacatttc ggactcccaa ggacgtacaa aatagaaaac
     6001 tatagaaaaa aataatcaga aaatagcatg tcatctcttt gtaaaacgcg tttgcaagaa
     6061 gaaaggaaac aatggagaag agatcatcca tttgtatgta aattttagta aacttgaaga
     6121 aatcactaac aacttctctt acttagggat tctatgcaaa accttgtaaa tcatctgatg
     6181 gaggactcga tttaatgaat tggaaggttg gatttccagg tggcactttt cggggaaatg
     6241 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6301 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6361 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6421 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6481 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6541 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6601 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6661 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6721 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6781 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6841 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6901 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6961 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7021 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggatctcgc ggtatcattg
     7081 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7141 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7201 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7261 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7321 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7381 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7441 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7501 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     7561 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7621 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7681 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7741 acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga
     7801 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7861 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7921 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7981 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8041 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8101 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
