LOCUS       sequence-263771-        8075 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression in fission yeast from promoter pnmt41 , integrated at 
            ura4 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8075)
  AUTHORS   Vjestica A, Marek M, Nkosi PJ, Merlini L, Liu G, Berard M, 
            Billault-Chaumartin I, Martin SG
  TITLE     A toolbox of stable integration vectors in the fission yeast 
            Schizosaccharomyces pombe.
  JOURNAL   J Cell Sci. 2020 Jan 8;133(1). pii: jcs.240754. doi: 
            10.1242/jcs.240754.
  PUBMED    31801797
REFERENCE   2  (bases 1 to 8075)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2020-01-8"; volume: "133(1). pii: jcs.240754. doi"; 
            pages: " 10.1242/jcs.240754"
COMMENT     Sequence Label: pAV0752
FEATURES             Location/Qualifiers
     source          1..8075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             1640..2434
                     /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      complement(2926..3381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3063..3084)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3275..3294
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3507..3529
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3521..3538
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3522..3538
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3548..3567
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3548..3566
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3608..4767
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     terminator      5541..5790
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5557..5575)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5815..5835)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5815..5833)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5854..5870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5854..5870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5867..5889)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5878..5894
                     /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)."
     promoter        complement(join(5902..5908,5909..5926,5927..5932))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5947..5968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6085..6102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6256..6844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6336..6355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7015..7806,7807..7875))
                     /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     7638..7657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7876..7980)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8048..8066
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaccg ggtattagtc aaaactagta tactttttct cggagtataa tacacaatat
      241 cggtgcaaat aggttttaaa attgctccaa tcacatgttc ttaagaaaaa aacgtcaaaa
      301 gaaatctaag tattcccaaa ataattcctc ctgacatgag aagaataaaa aacactaaga
      361 tgcattgtca gtatgaaaca caagccaaag ttgctaacaa taggatcaat tcttacttac
      421 tggcaatgtt acgaggaatc ttggtaggag gatgtttggc aacataacgt tcttttgaat
      481 ttgtttttac acgacgagta atgttcttct gaaactgtgt gtttgcacga cgctgagagg
      541 gagtttgaac cgtctaaaag acatagttag gatccatgat gcacagattt ctaaagataa
      601 aactacttac aggcatgttg gtttattaaa aaagaaaaac aggaatgata agagggaaag
      661 aatacgtgta cagagcgctt acataatgag tgagtgtcat tattgaggaa tgccatgtca
      721 gatttgacac aactcagaac ttattaactt tttatgattg gaacttttgt tttgcccaag
      781 gaatttttaa tggaagaagt tgttttataa atgatattga atttatatcc actcttttta
      841 gcgcatttgc atctctactt taggttcttc tttgactaga ttctaggatg cgtattggat
      901 atactggtct ttgaatcatt gtcgaaaaaa aaattcatgg ttgtacccat gagcaaactg
      961 cttttgtggt tatccatgtg cttcttttgt tcagcatcta tcttcttaat cgcatggaag
     1021 atttgattta aaccaaaaga aatattgtta aactttgaca gaagctttaa attttgattc
     1081 catacattaa atggataaaa ctcctgaagc ttagctacaa atcccactgg ctatatgtat
     1141 gcatttgtgt taaaaaagtt tgtatagatt atttaatcta ctcagcattc tttctctaaa
     1201 taggaatttg ttacttaatg gagaaaaaaa tgtttcgatt tacctagtgt atttgtttgt
     1261 atactcacgt ttaatttcaa acatccattc tatcttgtgt aatttttggc atggtgaaaa
     1321 agataatcag ccttataatc tttacaaaag taagaaattc tgtaaataag ccttaatgcc
     1381 cttgctttaa attaaaatgg ttctttttca tgataatgtt tgcactttgt gaatatattt
     1441 tagatagttc tgtgaggtat aattaagatg ttttagagac ttatacaatt ttgtctttat
     1501 aaattcttaa ttgattttac catcccagtt taactatgct tcgtcggcat ctctgcacat
     1561 gtcgtgtttt cttaccgtat tgtcctacca agaacctctt ttttgcttgg atcgaaatta
     1621 aaggtttaaa agcaaagtta tggatgctag agtatttcaa agctattcag ctagagctga
     1681 ggggatgaaa aatcccattg ccaaggaatt gttggctttg atggaagaaa agcaaagcaa
     1741 cttgtcagtc gcggtcgatt tgacgaagaa atccgaaatc ttagaattgg tagataaaat
     1801 tggaccctat gtctgtgtta tcaagacaca tattgacgtt gtcgaggatt tcgaccagga
     1861 tatggtagaa aaactggtgg ccttaggtaa aaagcatcgt tttcttatct ttgaggatcg
     1921 caaattcgca gacattggaa ataccgtcaa gctacaatat gcatctggtg tgtacaaaat
     1981 tgcttcttgg gctcatatca caaattgcca tacagtgcca ggcgagggta ttatacaagg
     2041 cctcaaagaa gttggtttac ctttgggacg tggtctcttg cttttggctg aaatgtcttc
     2101 caaaggctct ttggctactg gttcctacac agagaaaacc ttagaatggt ttgagaagca
     2161 taccgatttt tgctttggct ttatagctgg tcgtcgattt cctaaccttc aaagcgacta
     2221 cataactatg tcccctggta tcggcttgga tgttaaagga gacgggctgg gacagcaata
     2281 tcgtactcct gaagaagtga ttgtaaactg cggtagcgat atcatcattg ttggtcgtgg
     2341 agtttatgga gctggtcgta atcctgttgt cgaagccaag agatatagag aagctggttg
     2401 gaaggcatat cagcaaagac tttctcagca ttaaaaaaag actaatgtaa aatttttttg
     2461 gttggttatt gaaaaagtcg atgccttgtt tgcgtttgtt ttcctaggcg ttttatgtca
     2521 gaaggcattt agaattagta tacaagtact ctttggtaaa attttatgta gcgactaaaa
     2581 tattaactat tatagataaa caccttggga ataaaaagta atttgctata gtaatttatt
     2641 aaacatgctc ctacaacatt accacaatct tttctcttgg attgacattg aataagaaaa
     2701 gagtgaattt ttttagactt gtaatgataa ctatgtacaa agccaatgaa agatgtatgt
     2761 agatgaatgt aaaataccat gtagacaaac aagataaaac ttggttataa acattggtgt
     2821 tggaacagaa taaattagat gtcaaaaagt ttcgtcaata tcacaagctt gctagggcgg
     2881 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta
     2941 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3001 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3061 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3121 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3181 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3241 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3301 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3361 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3421 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3481 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3541 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggcccattt aaatgatcag
     3601 aaaattatcg ccataaaaga cagaataagt catcagcggt tgtttcattt cctatatttt
     3661 ttttttattt ttttattttt taataaggga aaatttaacg tctaaggata cagaagattg
     3721 ttagcacatt aaagtaataa aggcttaagt agtaagtgcc ttagcatgtt attgtatttc
     3781 aaaggacata atctaaaata ataacaatat catttctcac aagttattca attttctttt
     3841 ttttttctaa taatatcaag aatgtattat ttgtttgaca taagtcaact aatttattta
     3901 atatgctgga ttaatcttgc agacatgtaa attaacaagt tttagtcaaa taacgttgaa
     3961 gtttcaatga actcaaataa tttctctttt tttttatata accatatgtc taatctgatt
     4021 tatattttcc gcagggatca actgaagtta tgacatttgg attggatcac ttataacctt
     4081 ggtcgccaaa taatacaaaa atcagcgtta taaaacaaag aaggtttttg ttaagaaatt
     4141 aatcctcttt cttgataaga aagttgaacc gaaattgcag atactgatat atgaaaataa
     4201 tacccacaat tttgggaata gcgcaagcct caatttaaac aataggtgag gacacatgat
     4261 aatgacctca atgattgtta gaagaaaaga gcctcattac aaaatcgaaa aatgaatggt
     4321 tgggtacaag tttccaaaac atggtaaagt ggactttgcg tatgagacgt aaatagaaaa
     4381 aaacacttgt tatatgtttt ctagaattat tgttgtctct ttatggttgg atgatgcaaa
     4441 atagtaattt cggttagttg ctgtaaaaca ccacgagaca aatagatatg gatatttatt
     4501 aaatcaggaa aaacgtaact ctcggctact ggatggttca gtcacccaac gattactggg
     4561 gagagaaaac agggcaaaag caaagcttaa aggaatccga ttgtcattcg gcaatgtgca
     4621 gcgaaactaa aaaccggata atggacctgt taatcgaaac attgaagata aaggaagagg
     4681 aatcctggca tatcatcaat tgaataagtt gaattaatta tttcaatctc attctcactt
     4741 tctgacttat agtcgctttg ttaaatcctc gaggtcgacg aattcttaat taacatggca
     4801 agctccaagg gtgaagagct atttactggg gttgtaccca ttttggtaga actggacgga
     4861 gatgtaaacg gacataaatt ctctgttaga ggtgagggcg aaggcgatgc caccaatggt
     4921 aaattgactc tgaagtttat atgcactacg ggtaaattac ctgttccttg gccaacccta
     4981 gtaacaactt tgacatatgg tgttcaatgt ttctcaagat acccagacca tatgaaaagg
     5041 catgatttct ttaaaagtgc tatgccagaa ggctacgtgc aagagagaac tatctccttt
     5101 aaggatgacg gtacgtataa aacacgagca gaagtgaaat tcgaagggga tacactagtt
     5161 aatcgcatcg aattaaaggg tatagacttt aaggaagatg gtaatattct cggccataaa
     5221 cttgagtata atttcaactc gcataatgtg tacattacag ctgacaaaca aaagaacgga
     5281 attaaagcga attttaaaat caggcacaac gtcgaagatg ggtctgttca acttgccgat
     5341 cattatcagc aaaacacccc tattggtgat ggtccagtct tgttacccga taatcactac
     5401 ttaagcacac agtctagatt gtcaaaagat ccgaatgaaa agcgtgatca catggtttta
     5461 ttggaatttg tcaccgctgc aggaataact cacggaatgg acgagcttta taagcccggg
     5521 gcggccgctt aaggcgcgcc tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     5581 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     5641 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     5701 ttttttctgt acaaacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     5761 gttttgggac gctcgaaggc tttaatttgc agatctgagc tccagctttt gttcccttta
     5821 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5881 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5941 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6061 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7981 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     8041 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
