LOCUS       sequence-182949-       10039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Leu1 integrating plasmid with cdc25 genomic fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10039)
  AUTHORS   Keifenheim D, Sun XM, D'Souza E, Ohira MJ, Magner M, Mayhew MB, 
            Marguerat S, Rhind N
  TITLE     Size-Dependent Expression of the Mitotic Activator Cdc25 Suggests a 
            Mechanism of Size Control in Fission Yeast.
  JOURNAL   Curr Biol. 2017 May 22;27(10):1491-1497.e4. doi: 
            10.1016/j.cub.2017.04.016. Epub 2017 May 4.
  PUBMED    28479325
REFERENCE   2  (bases 1 to 10039)
  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"; doi: 
            "10.1016/j.cub.2017.04.016"; journalName: "Curr Biol"; date: 
            "2017-05-22- 22"; volume: "27"; issue: "10"; pages: "1491-1497.e4"
COMMENT     Sequence Label: pFS485
FEATURES             Location/Qualifiers
     source          1..10039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1537..1992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1674..1695)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1886..1905
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2118..2140
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2132..2149
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2133..2149
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2159..2178
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2159..2177
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7012..7032)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7012..7030)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7051..7067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7051..7067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7064..7086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7075..7091
                     /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(7099..7105,7106..7123,7124..7129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7144..7165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7282..7299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7453..8041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7533..7552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8212..9003,9004..9072))
                     /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     8835..8854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9073..9177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9245..9263
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9301..9323)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     9423..9442
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgctatggac acttggcctt acagtttgga agaagtttct cgtattgctc gtttggctgc
       61 ttggttagct gaaacttcca accctcctgc tcccgtcaca ttactcgaca aagctaatgt
      121 tttggcaact tctcgtcttt ggcgcaagac tgtagctaag atttttaagg aggaatatcc
      181 tcaccttacc ttaaaaaacc agctcattga ctccgcagcc atgcttttgg tcaagagccc
      241 tcgtacactt aacggtgttg ttttgactga caacttgttt ggtgacatta tttcagatga
      301 ggcttctgtc attcctggta gcttgggcct tttgccttct gcctcccttt ccggtgtggt
      361 aggaaaatca gaagaaaagg ttcattgttt ggttgagccc attcacggta gcgctcccga
      421 tatcgctggc aagggcattg ttaatcctgt tggtacaatt ttatctgctt cccttctcct
      481 tcgttatggt ttgaatgctc ctaaggaggc tgaagctatc gaagccgccg tacgcaaggt
      541 cttggatgat acttcaattg gtggacgtgg tctttatact cgcgatttgg gaggtgaggc
      601 ttctaccgct gatattacta aggctgttgt tgaagaactt gaaaaaattt tgtaataggg
      661 tttaatgtag aataaattca tatgatttga tttgacctaa tttttattga ttaattgtgt
      721 cgcatattta ttatcgttaa aaatgcaatt gacaaaagcg taaattttaa ggctctaatg
      781 ttttttcttt tggtatgtta tagtcaagca aagatgtatt aagttgcaaa agcatggaat
      841 tgatttacga catccgttca actttatgac cgagttgtcg ctgtgtactt actctttttt
      901 tactatgaaa agtttcaaac ataatatcat taattatttc tttttaaacg aagtaacctg
      961 taacataaat aataggggaa aagaagaaaa aaaccataaa aattcgagaa cgtaacaatt
     1021 attattgtaa cgatgacaac ccgattcttt gagagagcca gagacatgac ttttgcatta
     1081 ccgtaattgg tttctactta tgaattgtaa acttgtgtag ggaatttcat ccacagctat
     1141 cgcgggctgc tctctgcatc agtgttagca cattttatat taatgaagat tttgctatgc
     1201 aaaatacgta catttctggc gcatatattt ttatcaaacg aacatgacta tcctttccac
     1261 tttgatggtt cgcaagtatt ttgatttgta ttaaatgcac atctaacaag ccatttcgcc
     1321 caaaatcacc aactggcaca tgtacccaac cagttagttc ctcgacttga actgttgtaa
     1381 caatttccaa atcttgaaac cctgtcccag ctgatattcg caaagtactg ggagtataag
     1441 attcatcaag ggtgtattga aggtacatac tcacatactt aatcgatgcg gtgtgaaata
     1501 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
     1561 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1621 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1681 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1741 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1801 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1861 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1921 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1981 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     2041 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     2101 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     2161 atacgactca ctatagggcg aattgggtac cggggaagca tggtcgttat tcataaagat
     2221 accaaaatct ttaaattagt gtacctaaaa agatttgttt gtttgaacat taagtgctct
     2281 gcaaactttc atacttaaac aattcgttat ttatgattac gaggtttatc ctacaatgta
     2341 ttaaatacaa tcgttggcgt tcaaagctgc acttcatagg gcgttgatgt cataatcaat
     2401 aaaattcata taagcgaatt taaggttgac ttagatgacg acattctgat tgttgcaaat
     2461 aaaaagtaaa caaaacaaaa tctacaaata tcttcaaatt ttaaaaaagc acacatatga
     2521 ttcactttct caacggaaca tgtttaaatt tttaccaaaa aagtagtatt gattaaaaat
     2581 tttcgtaatt ttttttttgg gaaggtactt aagtgtgaat gtccaacttc cgcattgtaa
     2641 ctattttata tcacataaaa cttcgtaact ctctggtgaa cttaaccttc cattggaaag
     2701 ggtaagctac aatactaccg cttacagaac atgcataaaa ggaagtgatt taaaaataaa
     2761 agttttattc gttttctact taaatatttc catcagagat ggttaggtag agttacaaaa
     2821 acgtaagcaa ctcagttcct atcgcattac tgcttcaatc aaagaatgct aaattcttag
     2881 cattcctatc aacagcaatt gcgagaagag caattagaat ggacttcggt acgtatgttt
     2941 acccgtgatt gcttagaaat tattaagctc aattgattat aaaataatca attattaaac
     3001 caaaatccat gcgtaataat aataaccaaa ctagtcgttc tagatattta gtcgtaaaag
     3061 tgatgtactt cgaaaagttt agccagttta aattttttgt tgtcgagacc ttttaaaagg
     3121 tatattgctt tctttcaata ataataataa aaaatagaaa taaaaggttt catgattcaa
     3181 agtttataga aaaaacttag gtttagaaag ttgaatatat aagagtatac ttcaggctag
     3241 gtaaagtatt gagtcagcct aaaatcatta aaatcttcta agtgtagaga gggaatgcat
     3301 ggcagtagga gaatcattaa cgtctgggga agctaacaca ctttggccaa aagtataact
     3361 tttagtacgc ataaaagtag cgtttcgttt gaaattattc atagccttgg tgcaggtcat
     3421 aacatgcgaa gcatcgttca tcggaacgta gttaattggg tcacaacggt ttttgtggtt
     3481 ttcgtaaaac gacttgtaac caccatgaag tatataaacc tcgggatagt aaaggaaagg
     3541 ataacgatga ctattcattc gtctgtcggt atttcgaaag tgcaatgcca aatgaggtgc
     3601 tctgtgcgca ctatgttcac aatgaaaaac aagtgcaacc cgatgagtaa gaggtttact
     3661 taaaaaggca tccacaatag cttgtttcgt atttaaattc acagcagtgc taatatggcc
     3721 cccaaggtat tcatattcaa aacggcaatc aataatgatg catttgtcaa aaatatcctt
     3781 aaatttacca tctaatagac cgagcaatgt ttcttgggta attcgtttca atgagtcctc
     3841 cttcacggcg aagcaaggaa gagataaatc ctcgacatga gaagagatga agcgctcact
     3901 ttctttggtc gattgttttg gcgtaacgca cacaagatct tggcttttga aaagccctag
     3961 tcttgtggaa ttgagaaaca tgctttgggt acgacgaagc actggtgtat cttgctcatc
     4021 atattccgca tcttcttgca tagcgaatgc catcggagaa ggggacctaa ctttcaagct
     4081 aggaaaggca atatctttat ttatcttggg taacatatca ataggtttaa tgggtgaagc
     4141 ttgaaaaaga tcatctaaag aattatcatt gcaagcacca aaatcactgt tgggacgttg
     4201 aatagaggca gcgtcttcaa caagttcaac agagctttct ggtaaatcag caaccttgcc
     4261 catcgtaaat gtatcctcat gatcagacat taattcgaaa tcatccgtat catcagtaag
     4321 acaactttca agtaagctgg tcgtgttgct cgactgacta caggtacggg ataaggcata
     4381 agtcaaatgg cgggtagctt gtccaccaga agttcccttc tttttgttaa tggaatatga
     4441 ggaactcgat ctggatcgca aaaatggggg tgcgttgcct gatgatcgag aacgtgagac
     4501 gttaggacga aggtaggaat caaaagaaga actcgataat gacgaacgag tagcttcgct
     4561 tttggtggtg tttaatgcga ttgatgcttt aggcttaaag ctgtaagcaa tagtgctgga
     4621 aacacgtggg gaatcttggg taatggaata cttcgatgca ggttgccgga agaagtagga
     4681 ctcggttaaa gcgtcattcg agggtgactc cgggagaggt gaaacaatag tcttgttagc
     4741 gagaggggtt tctacagtac aagaaagaga tcgaaagagc gtacgacgag gagtcggtaa
     4801 ggcagggctt tcatcgatat gcatgttcat tgacatatca gcagctaagg aagatgcagg
     4861 tgaagctgtc gaacaagttt ttccaaaagg atccactaat gttgaggcaa tgtgtttgga
     4921 tttcgggaag aaaaaatcca gctcttgatt tgcattgcga tccgacatca actttaattc
     4981 acgggcagca ggacgcaata cattccgttt gccagataga gtgttggtaa aggaaagtga
     5041 agaaagcgga gaatccattt tagttgaggt ttaactatta ttaaaagcag acaaaatggt
     5101 aaatttaatt taaaccaagt acactgcaaa aagaactaat gggacgacac gactgcaaaa
     5161 gattgagttt cagaatgaga aatcaataga gcaagtaaga taaattataa aggaaaatcc
     5221 tagggaagag tcaaaagacc ggtttaagat attaatgcca aagtgagaca aaaagaaaac
     5281 taaaagaaag acaggatgaa agcaatatag ccaaatatta aacaaaacca acctttacaa
     5341 aagaaaccgg aaagaagttt aagtaagttt agactccaca cgacgggaag accgaataaa
     5401 agtatataaa aaaaaagaga ttaatgagac aaaaatagat gagcaccaac ggtcacgact
     5461 ataatataga atataaaaga tactgtcgtt gaaagaaaaa atgaaagaaa aaagaaagtt
     5521 aaaagaaagc agttgattaa tatgaaaaca acagaatacc caaaagttca gactcaaagt
     5581 tcgttacaag taataaaccc cccactttac catataaagg cttcttgaac aaataacttc
     5641 aagaatccta ttaattgcct taaaaagcgt aagcaattgt attatagacg tttcaaaaaa
     5701 tcactgaacc agcagttagt acttgagaag catcaacagg aatccaactt gaggagagaa
     5761 ggatgcccga aactaacagg catcaagttc tttttaaact tacaactttg gtattcttaa
     5821 gaataatatt ttcaatgtaa tacacttgat ctaaaggaga agagcttgaa gcgctgtaga
     5881 tagttgaggg tagagagaaa tctcagctct gcctccgaga ggaaaaaaaa gcaaaggtaa
     5941 caacaacaat aaaacaccaa ccaacaaatt cgagtaacag aaaacaataa taaaaacaaa
     6001 tacatgatta aagcaaagtt aaactgtaag ctgtaatact tagccaacac ctcagccatg
     6061 acaactattg ttgtatagtt tagttgcgta gataatgccg tttcaagcta caatatgtca
     6121 cctcttaatc tattaattat ataaaagcaa ataatcattt cattaacaac attgaactag
     6181 atgaaaacac ataaattttt gcatcttgta tgtttgttta catgtccata catctcagct
     6241 attcgatcca ccgcaggttt gtctgatttg ttgccctagc aacgccaaat gttgtgtatg
     6301 tgtaaacaaa tacataagaa caaacgtcag gtaacctgct tgacagccat taaatcccca
     6361 taatctcata cccgcactcc tgatatgtaa cacgagcagc gagataaaat atgttatcag
     6421 gtgttagcga ataatcgtaa cacgatgaat ctctctactt cgggactcgg actgtcattc
     6481 cgaattggaa aacgagggtg taactcattg aaagtctccc gatgttatgt agctatgggt
     6541 tgataccatt attgtcgtag taggcatatt tcgcaagata gtatagccag ttgtcttgtt
     6601 aaactcggat atcttcaaca aagataccgc tattattaca cttcaacata caaaatataa
     6661 caaaactcta acctttattt aaagtattct agcaatcaga ggcagatatc gtatccacag
     6721 aaacactaga gcaaaaaaga aaatatttta tcaggtggag cacctttttg taaaaacgcc
     6781 ctgatacgtg attagtagac aaagaaaagt gtaaccataa aaattgctgg aacaaattgt
     6841 acgcaaaaag atggagtatg aaacaaataa aaagtgcagt ataaaacagt ttgtcgaaca
     6901 cttcgtggag ttcagtgaca caaaatcaaa tgagtatccg taaaacctaa aaacaatgag
     6961 tggacttaat tcttaagatg ctcggcaaga atggagctcc agcttttgtt ccctttagtg
     7021 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7201 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7261 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7321 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7381 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8101 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8161 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8221 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8281 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8341 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8401 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8461 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8521 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8581 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8641 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8701 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8761 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8821 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8881 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8941 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9001 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9061 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9121 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9181 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     9241 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     9301 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     9361 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     9421 tgcggcatca gagcagattg tactgagagt gcaccatacg atatcccaat ctgtagtata
     9481 aaaagtgcaa cagtgttgag tttcccgaaa ccaggcattt ttgcggtagg gtctgaaact
     9541 ttcacgaaac catttcacaa caaaacttct tcaacaacat gtgtgcaaag aaaatcgtcg
     9601 tcttaccagg agaccatatt ggccctgaaa ttgttgcttc tgccttggag gttttgaaag
     9661 tcgttgagaa gaagcgacct gagttaaaac tcgagtttga agaacacaag attggaggtg
     9721 cctctattga tgcctatgga acccctttga ctgacgagac tgtgaaggct tgtttggaag
     9781 ctgacggtgt tcttttgggt gccgttggtg gtcctgaatg gaccaacccc aattgtcgtc
     9841 ctgagcaagg tttattgaag cttcgtaaga gtatgggtgt ttgggccaac cttcgacctt
     9901 gcaactttgc cagcaagtct ttagtcaagt acagcccttt gaagcctgaa atcgttgaag
     9961 gtgtcgattt ttgtgttgta cgagaactta ctggaggttg ttactttggt gagcgcactg
    10021 aggacaacgg atcgggtta
//
