LOCUS       sequence-337157-        6512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid containing the ura4 genomic region,which flanked by a NotI
            site at the both ends. It served as the control plasmid or backbone 
            for the ura4 integrated plasmid construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6512)
  AUTHORS   Zhang XR, Zhao L, Suo F, Gao Y, Wu Q, Qi X, Du LL
  TITLE     An improved auxin-inducible degron system for fission yeast.
  JOURNAL   G3 (Bethesda). 2022 Jan 4;12(1):jkab393. doi: 
            10.1093/g3journal/jkab393.
  PUBMED    34849776
REFERENCE   2  (bases 1 to 6512)
  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: "G3 
            (Bethesda)."; date: "2022-01-4"; volume: "12(1):jkab393. doi"; 
            pages: " 10.1093/g3journal/jkab393"
COMMENT     Sequence Label: pDB4697
FEATURES             Location/Qualifiers
     source          1..6512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     210..229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     463..480
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    597..618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(633..638,639..656,657..663)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    671..687
                     /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)."
     primer_bind     676..698
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     695..711
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     695..711
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      787..796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1614..2408)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRQFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     primer_bind     complement(3435..3454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3436..3454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3461..3478)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3461..3477)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3470..3492)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3618..4046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3705..3724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3915..3936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             4390..5184
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4444..4463)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5054..5073
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(5202..5270,5271..6062)
                     /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     complement(5420..5439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6233..6512,1..309)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
       61 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
      121 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      181 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      241 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      301 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      361 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
      421 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      481 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      541 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      601 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      661 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      721 gccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattgcc
      781 cttgcggccg ccatggatcc taactatgtc ttttagacgg ttcaaactcc ctctcagcgt
      841 cgtgcaaaca cacagtttca gaagaacatt actcgtcgtg taaaaacaaa ttcaaaagaa
      901 cgttatgttg ccaaacatcc tcctaccaag attcctcgta acattgccag taagtaagaa
      961 ttgatcctat tgttagcaac tttggcttgt gtttcatact gacaatgcat cttagtgttt
     1021 tttattcttc tcatgtcagg aggaattatt ttgggaatac ttagatttct tttgacgttt
     1081 ttttcttaag aacatgtgat tggagcaatt ttaaaaccta tttgcaccga tattgtgtat
     1141 tatactccga gaaaaagtat actagttttg aataataaag cttgtgatat tgacgaaact
     1201 ttttgacatc taatttattc tgttccaaca ccaatgttta taaccaagtt ttatcttgtt
     1261 tgtctacatg gtattttaca ttcatctaca tacatctttc attggctttg tacatagtta
     1321 tcattacaag tctaaaaaaa ttcactcttt tcttattcaa tgtcaatcca agagaaaaga
     1381 ttgtggtaat gttgtaggag catgtttaat aaattactat agcaaattac tttttattcc
     1441 caaggtgttt atctataata gttaatattt tagtcgctac ataaaatttt accaaagagt
     1501 acttgtatac taattctaaa tgccttctga cataaaacgc ctaggaaaac aaacgcaaac
     1561 aaggcatcga ctttttcaat aaccaaccaa aaaaatttta cattagtctt tttttaatgc
     1621 tgagaaagtc tttgctgata tgccttccaa ccagcttctc tatatctctt ggcttcgaca
     1681 acaggattac gaccagctcc atagactcca cgaccaacaa tgatgatatc gctaccgcag
     1741 tttacaatca cttcttcagg agtacgatat tgctgtccca gcccgtctcc tttaacatcc
     1801 aagccgatac caggggacat agttatgtag tcgctttgaa ggttaggaaa ttgacgacca
     1861 gctataaagc caaagcaaaa atcggtatgc ttctcaaacc attctaaggt tttctctgtg
     1921 taggaaccag tagccaaaga gcctttggaa gacatttcag ccaaaagcaa gagaccacgt
     1981 cccaaaggta aaccaacttc tttgaggcct tgtataatac cctcgcctgg cactgtatgg
     2041 caatttgtga tatgagccca agaagcaatt ttgtacacac cagatgcata ttgtagcttg
     2101 acggtatttc caatgtctgc gaatttgcga tcctcaaaga taagaaaacg atgcttttta
     2161 cctaaggcca ccagtttttc taccatatcc tggtcgaaat cctcgacaac gtcaatatgt
     2221 gtcttgataa cacagacata gggtccaatt ttatctacca attctaagat ttcggatttc
     2281 ttcgtcaaat cgaccgcgac tgacaagttg ctttgctttt cttccatcaa agccaacaat
     2341 tccttggcaa tgggattttt catcccctca gctctagctg aatagctttg aaatactcta
     2401 gcatccataa ctttgctttt aaacctttaa tttcgatccg agcaaaaaag aggttcttgg
     2461 taggacaata cggtaagaaa acacgacatg tgcagagatg ccgacgaagc atagttaaac
     2521 tgggatggta aaatcaatta agaatttata aagacaaaat tgtataagtc tctaaaacat
     2581 cttaattata cctcacagaa ctatctaaaa tatattcaca aagtgcaaac attatcatga
     2641 aaaagaacca ttttaattta aagcaagggc attaaggctt atttacagaa tttcttactt
     2701 ttgtaaagat tataaggctg attatctttt tcaccatgcc aaaaattaca caagatagaa
     2761 tggatgtttg aaattaaacg tgagtataca aacaaataca ctaggtaaat cgaaacattt
     2821 ttttctccat taagtaacaa attcctattt agagaaagaa tgctgagtag agtaaataat
     2881 ctatacaaac ttttttaaca caaatgcata catatagcca gtgggatttg tagctaagct
     2941 tcaggagttt tatccattta atgtatggaa tcaaaattta aagcttctgt caaagtttaa
     3001 caatatttct tttggtttaa atcaaatctt ccatgcgatt aagaagatag atgctgaaca
     3061 aaagaagcac atggataacc acaaaagcag tttgctcatg ggtacaacca tgaatttttt
     3121 tttcgacaat gattcaaaga ccagtatatc caatacgcat cctagaatct agtcaaagaa
     3181 gaacctaaag tagagatgca aatgcgctaa aaagagtgga tataaattca atatcattta
     3241 taaaacaact tcttccatta aaaattcctt gggcaaaaca aaagttccaa tcataaaaag
     3301 ttaataagtt ctgagttgtg tcaaatctga catggcattc ctcaataatg acactcacgc
     3361 ggccgcaagg gcaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     3421 gagggcccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     3481 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3541 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3601 gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3661 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3721 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3781 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3841 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3901 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3961 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4021 gatttaacaa aaatttaacg cgaattttaa caaaattcag ggcgcaaggg ctgctaaagg
     4081 aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     4141 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     4201 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     4261 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     4321 ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga
     4381 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4441 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4501 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4561 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4621 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4681 ccggggcagg atctcctgtc atcccacctt gctcctgccg agaaagtatc catcatggct
     4741 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4801 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4861 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4921 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4981 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5041 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5101 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5161 cgccttcttg acgagttctt ctgaattgaa aaaggaagag tatgagtatt caacatttcc
     5221 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5281 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5341 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5401 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5461 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5521 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5581 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5641 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5701 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5761 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5821 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5881 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5941 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6001 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6061 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6121 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6181 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6241 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6301 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6361 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6421 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6481 gcgataagtc gtgtcttacc gggttggact ca
//
