LOCUS       sequence-263773-        7590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in fission yeast from promoter ppom1 , integrated at ura4
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7590)
  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 7590)
  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: "J Cell 
            Sci."; date: "2020-01-8"; volume: "133(1). pii: jcs.240754. doi"; 
            pages: " 10.1242/jcs.240754"
COMMENT     Sequence Label: pAV0747
FEATURES             Location/Qualifiers
     source          1..7590
                     /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"
     terminator      5056..5305
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5072..5090)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5330..5350)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5330..5348)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5369..5385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5369..5385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5382..5404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5393..5409
                     /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(5417..5423,5424..5441,5442..5447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5462..5483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5600..5617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5771..6359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5851..5870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6530..7321,7322..7390))
                     /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     7153..7172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7391..7495)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7563..7581
                     /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 aaatagagat
     3601 gtggtctatc catcgaaaag attattaaag aagatttagt aaattagtcg tgtatttcgt
     3661 atttaatcag cttaaaacac tgtatataag cgcaatttta tgtctgttca gcgagcaatt
     3721 taatttgttc tgcaaaaagg aagcctttac tgccatcaaa ctatttgcta gaaacgcatt
     3781 aattgcaaaa acaagtaggt aattttgaac cgcctaaaac ttagtttcaa tgggaaggtc
     3841 tgtgtgattg atttatcttt ataacgaggg ttaaaacagt agttatgggg aattttatat
     3901 gtatacaaat gaaaaccaat tttaaacctt ggactaaaaa aaacccaaag tatgttcact
     3961 caaactcttt ttaaagcatc aaaatatata tgcagtaaat cttaataaaa tattttcttt
     4021 ttggtaaaat ttgtgcacaa atcaataagc gttctatgtt tcatttacag cttgcgtgcc
     4081 ctttcagaag tggttggtat tgatatgcag ggtgtagcgc ggctttaatt agccacgaac
     4141 gtactgctac tagtttgcat tttaaagaag ataagagact gtaactgaag gagggagctt
     4201 atagaaacaa tattttgttt caaaaaagcc agtaaaagcg taatatagtg agaattttgg
     4261 ctttttgaag caaagtttgc aactcgaggt cgacgaattc ttaattaaca tggcaagctc
     4321 caagggtgaa gagctattta ctggggttgt acccattttg gtagaactgg acggagatgt
     4381 aaacggacat aaattctctg ttagaggtga gggcgaaggc gatgccacca atggtaaatt
     4441 gactctgaag tttatatgca ctacgggtaa attacctgtt ccttggccaa ccctagtaac
     4501 aactttgaca tatggtgttc aatgtttctc aagataccca gaccatatga aaaggcatga
     4561 tttctttaaa agtgctatgc cagaaggcta cgtgcaagag agaactatct cctttaagga
     4621 tgacggtacg tataaaacac gagcagaagt gaaattcgaa ggggatacac tagttaatcg
     4681 catcgaatta aagggtatag actttaagga agatggtaat attctcggcc ataaacttga
     4741 gtataatttc aactcgcata atgtgtacat tacagctgac aaacaaaaga acggaattaa
     4801 agcgaatttt aaaatcaggc acaacgtcga agatgggtct gttcaacttg ccgatcatta
     4861 tcagcaaaac acccctattg gtgatggtcc agtcttgtta cccgataatc actacttaag
     4921 cacacagtct agattgtcaa aagatccgaa tgaaaagcgt gatcacatgg ttttattgga
     4981 atttgtcacc gctgcaggaa taactcacgg aatggacgag ctttataagc ccggggcggc
     5041 cgcttaaggc gcgcctcatg taattagtta tgtcacgctt acattcacgc cctcctccca
     5101 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     5161 ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     5221 tctgtacaaa cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     5281 gggacgctcg aaggctttaa tttgcagatc tgagctccag cttttgttcc ctttagtgag
     5341 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5401 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5461 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5521 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5581 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5641 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5701 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5761 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5821 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5881 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5941 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6001 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6061 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6121 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6181 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6241 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6301 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6361 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6421 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6481 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6541 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6601 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6661 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6721 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6781 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6841 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6901 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6961 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7021 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7081 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7141 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7201 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7261 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7321 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7381 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7441 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7501 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     7561 aaaataggcg tatcacgagg ccctttcgtc
//
