LOCUS       sequence-266123-        7724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in fission yeast from promoter pact1, integrated at ura4 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7724)
  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 7724)
  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: pAV0714
FEATURES             Location/Qualifiers
     source          1..7724
                     /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      5190..5439
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5206..5224)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5464..5484)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5464..5482)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5503..5519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5503..5519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5516..5538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5527..5543
                     /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(5551..5557,5558..5575,5576..5581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5596..5617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5734..5751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5905..6493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5985..6004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6664..7455,7456..7524))
                     /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     7287..7306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7525..7629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7697..7715
                     /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 aaatgatcta
     3601 cgataatgag acggtgtttg ggttgttttt actgaatttt tcttttattt tatgacatct
     3661 taatttttat gttaatgatt attcttatac ttttatctat ctttgattaa aactaatatt
     3721 cattcttttg atcactaatt atttttatag tatacgattc aacccaacta taatacattg
     3781 tatcaagaga aatttttaat tagatgtaat ttaatagcca tttgataaaa aaaaattacc
     3841 tttttttttg ctagtgttaa attattgatc ataacaatgt cgaatgaata ctaataaaat
     3901 agggacacgc gagttgcaaa tacttgaaat acctttgaag atttgtctgt gaattttgat
     3961 aagattttta tgaccatctc ttttatattc gatatcgtga actttcattt tcttttgttt
     4021 tttgcttttc aagttaaata acgcggaaat tatatttcct tatatgttta taagagcttt
     4081 taaaggattt gctatatcgc ttacagtact acaaccggtt gctcaatgtt atccgtttcc
     4141 gcggtcttct tccgtgcgca ttctgccgtg aagtgctaac gctgtgtgtg gctttatcaa
     4201 aaaagtggct caaatatagc aatttcatac atatttaaac acccattata cttaaatgta
     4261 tattttaaaa tatataagag attttcactg tataactgtg tatcttatct atatgtaatc
     4321 aacggcttca taccacctca gccagccgtg ttataactta ccgtttacca actacatttt
     4381 ttgtaacgaa ccaaaaaacc ctcaaaagac aagaccctcg aggtcgacga attcttaatt
     4441 aacatggcaa gctccaaggg tgaagagcta tttactgggg ttgtacccat tttggtagaa
     4501 ctggacggag atgtaaacgg acataaattc tctgttagag gtgagggcga aggcgatgcc
     4561 accaatggta aattgactct gaagtttata tgcactacgg gtaaattacc tgttccttgg
     4621 ccaaccctag taacaacttt gacatatggt gttcaatgtt tctcaagata cccagaccat
     4681 atgaaaaggc atgatttctt taaaagtgct atgccagaag gctacgtgca agagagaact
     4741 atctccttta aggatgacgg tacgtataaa acacgagcag aagtgaaatt cgaaggggat
     4801 acactagtta atcgcatcga attaaagggt atagacttta aggaagatgg taatattctc
     4861 ggccataaac ttgagtataa tttcaactcg cataatgtgt acattacagc tgacaaacaa
     4921 aagaacggaa ttaaagcgaa ttttaaaatc aggcacaacg tcgaagatgg gtctgttcaa
     4981 cttgccgatc attatcagca aaacacccct attggtgatg gtccagtctt gttacccgat
     5041 aatcactact taagcacaca gtctagattg tcaaaagatc cgaatgaaaa gcgtgatcac
     5101 atggttttat tggaatttgt caccgctgca ggaataactc acggaatgga cgagctttat
     5161 aagcccgggg cggccgctta aggcgcgcct catgtaatta gttatgtcac gcttacattc
     5221 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     5281 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     5341 atttttcttt tttttctgta caaacgcgtg tacgcatgta acattatact gaaaaccttg
     5401 cttgagaagg ttttgggacg ctcgaaggct ttaatttgca gatctgagct ccagcttttg
     5461 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     5521 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5581 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5641 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5701 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5761 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5821 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5881 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5941 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6001 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6061 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6421 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6481 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6541 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6601 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6661 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6721 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6781 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6841 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6901 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6961 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7021 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7081 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7141 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7201 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7261 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7321 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7381 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7441 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7501 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7621 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7681 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
