LOCUS       sequence-263756-        7902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in fission yeast from promoter ptdh1 , integrated at ura4
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7902)
  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 7902)
  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: pAV0749
FEATURES             Location/Qualifiers
     source          1..7902
                     /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      5368..5617
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5384..5402)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5642..5662)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5642..5660)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5681..5697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5681..5697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5694..5716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5705..5721
                     /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(5729..5735,5736..5753,5754..5759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5774..5795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5912..5929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6083..6671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6163..6182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6842..7633,7634..7702))
                     /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     7465..7484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7703..7807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7875..7893
                     /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 aaattaaagt
     3601 atggaaaatc aaaattcctg aatttttgta aactcccata caaatacgat tacacataca
     3661 tacaaaaatg caaataacaa aatatacacg cacgcatgca cactaaataa acaaactgtg
     3721 cgtccattcc ttttatgcac aaacgttgaa gctcaggaca tcgcaccaac aatcactcat
     3781 ctcttctctc tctacccact ttccacccac acccacagtc tgcatctgtt tgacgactgt
     3841 agctcaagca tcagtagcag ccgctgtcat cctcaactgc tgctgctgat gctgcttgaa
     3901 tgtacgcatg atagcaacga cgcaccgctc taccactact accgaagcgc catcatcacc
     3961 tcctcaccta ccatcgcaat cgtcctctcg aaactcttcg accgctttca cccggcttcc
     4021 tgcaggtaac aatgcgctcg gcgcttgatt aaaagaggcc agaggcgata aatgactcag
     4081 ggtgcatttg gcctcgctac gaaaggaccc tgtcttgtga tttttccgta gccaatggac
     4141 aaccgccttg tcaacctttt gactttggaa tgtgctagca attcctcctt ccccctgact
     4201 tcaggtgaca aggcttgcat cacgctgagg taggcgtcgg aaaaactttc ctcacaaggc
     4261 tgacgtcatg gaatgtgcaa tcaaagtcag gagcgggaac gaggcagatt accaacgaag
     4321 caacgaccat ctcggaacaa cttgcaaaga aagtcaagga aagttggcac cacatcacga
     4381 tcgcgcacct tctccaatca ccttgctcct tagttgttcc gaatcgagtg cgaggcaatt
     4441 gtggtaaaga tagtcggggg aaacccttac cccattcaca ccacggctga tggctgctgt
     4501 gtgggatgga aaggtataaa tagaggcagg cggtgcctag catcgcttct gtatagatca
     4561 ttcatccata gtattgattt acacttgatt caaactcgag gtcgacgaat tcttaattaa
     4621 catggcaagc tccaagggtg aagagctatt tactggggtt gtacccattt tggtagaact
     4681 ggacggagat gtaaacggac ataaattctc tgttagaggt gagggcgaag gcgatgccac
     4741 caatggtaaa ttgactctga agtttatatg cactacgggt aaattacctg ttccttggcc
     4801 aaccctagta acaactttga catatggtgt tcaatgtttc tcaagatacc cagaccatat
     4861 gaaaaggcat gatttcttta aaagtgctat gccagaaggc tacgtgcaag agagaactat
     4921 ctcctttaag gatgacggta cgtataaaac acgagcagaa gtgaaattcg aaggggatac
     4981 actagttaat cgcatcgaat taaagggtat agactttaag gaagatggta atattctcgg
     5041 ccataaactt gagtataatt tcaactcgca taatgtgtac attacagctg acaaacaaaa
     5101 gaacggaatt aaagcgaatt ttaaaatcag gcacaacgtc gaagatgggt ctgttcaact
     5161 tgccgatcat tatcagcaaa acacccctat tggtgatggt ccagtcttgt tacccgataa
     5221 tcactactta agcacacagt ctagattgtc aaaagatccg aatgaaaagc gtgatcacat
     5281 ggttttattg gaatttgtca ccgctgcagg aataactcac ggaatggacg agctttataa
     5341 gcccggggcg gccgcttaag gcgcgcctca tgtaattagt tatgtcacgc ttacattcac
     5401 gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     5461 tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat
     5521 ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac attatactga aaaccttgct
     5581 tgagaaggtt ttgggacgct cgaaggcttt aatttgcaga tctgagctcc agcttttgtt
     5641 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5701 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5761 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5821 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5881 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5941 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6961 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7381 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7441 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7501 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7561 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7621 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7681 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7741 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7801 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7861 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
