LOCUS       sequence-428700-        7415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty integration vector for genomic integration for multigene into 
            S. pombe. Targets Ura4 locus. Part type 15678 following the 
            YeastToolkit MoClo grammar. Contains GFP drop out..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7415)
  AUTHORS   Hebra T, Smrckova H, Elkatmis B, Prevorovsky M, Pluskal T
  TITLE     POMBOX: A Fission Yeast Cloning Toolkit for Molecular and Synthetic 
            Biology.
  JOURNAL   ACS Synth Biol. 2023 Nov 22. doi: 10.1021/acssynbio.3c00529.
  PUBMED    37991801
REFERENCE   2  (bases 1 to 7415)
  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: "ACS Synth 
            Biol. 2023 Nov 22. doi: 10.1021/acssynbio.3c00529."
COMMENT     Sequence Label: pPOM042_P15678_MultiGeneBackbone
FEATURES             Location/Qualifiers
     source          1..7415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(13..804)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1234..1822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1723..1742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2899..3693
                     /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"
     RBS             4462..4473
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4480..5193
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5205..5276
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5292..5319
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     gene            5570..6925
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     CDS             5914..6723
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5981..6000)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6728..6925
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 ccgagcggcc gcttatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
       61 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
      121 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
      181 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
      241 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
      301 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
      361 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
      421 gtcgatcgtg gctggctcga agatacccgc aagaatgtca ttgcgctgcc attctccaaa
      481 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
      541 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
      601 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
      661 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
      721 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
      781 ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac tgccctgctg
      841 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
      901 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac
      961 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     1021 catacgctac ttgcattaca gcttacgaac cgaacaggct tatgtccact gggttcgtgc
     1081 cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggtagcagcg aagtcgaggc
     1141 atttctgtcc tggctggtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1201 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1261 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1321 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1381 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1441 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1501 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1561 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1621 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1681 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1741 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1801 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1861 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1921 tgcggccgcc aatatgctta cataatgagt gagtgtcatt attgaggaat gccatgtcag
     1981 atttgacaca actcagaact tattaacttt ttatgattgg aacttttgtt ttgcccaagg
     2041 aatttttaat ggaagaagtt gttttataaa tgatattgaa tttatatcca ctctttttag
     2101 cgcatttgca tctctacttt aggttcttct ttgactagat tctaggatgc gtattggata
     2161 tactggtctt tgaatcattg tcgaaaaaaa aattcatggt tgtacccatg agcaaactgc
     2221 ttttgtggtt atccatgtgc ttcttttgtt cagcatctat cttcttaatc gcatggaaga
     2281 tttgatttaa accaaaagaa atattgttaa actttgacag aagctttaaa ttttgattcc
     2341 atacattaaa tggataaaac tcctgaagct tagctacaaa tcccactggc tatatgtatg
     2401 catttgtgtt aaaaaagttt gtatagatta tttaatctac tcagcattct ttctctaaat
     2461 aggaatttgt tacttaatgg agaaaaaaat gtttcgattt acctagtgta tttgtttgta
     2521 tactcacgtt taatttcaaa catccattct atcttgtgta atttttggca tggtgaaaaa
     2581 gataatcagc cttataatct ttacaaaagt aagaaattct gtaaataagc cttaatgccc
     2641 ttgctttaaa ttaaaatggt tctttttcat gataatgttt gcactttgtg aatatatttt
     2701 agatagttct gtgaggtata attaagatgt tttagagact tatacaattt tgtctttata
     2761 aattcttaat tgattttacc atcccagttt aactatgctt cgtcggcatc tctgcacatg
     2821 tcgtgttttc ttaccgtatt gtcctaccaa gaacctcttt tttgcttgga tcgaaattaa
     2881 aggtttaaaa gcaaagttat ggatgctaga gtatttcaaa gctattcagc tagagctgag
     2941 gggatgaaaa atcccattgc caaggaattg ttggctttga tggaagaaaa gcaaagcaac
     3001 ttgtcagtcg cggtcgattt gacgaagaaa tccgaaatct tagaattggt agataaaatt
     3061 ggaccctatg tctgtgttat caagacacat attgacgttg tcgaggattt cgaccaggat
     3121 atggtagaaa aactggtggc cttaggtaaa aagcatcgtt ttcttatctt tgaggatcgc
     3181 aaattcgcag acattggaaa taccgtcaag ctacaatatg catctggtgt gtacaaaatt
     3241 gcttcttggg ctcatatcac aaattgccat acagtgccag gcgagggtat tatacaaggc
     3301 ctcaaagaag ttggtttacc tttgggacgt ggactcttgc ttttggctga aatgtcttcc
     3361 aaaggctctt tggctactgg ttcctacaca gagaaaacct tagaatggtt tgagaagcat
     3421 accgattttt gctttggctt tatagctggt cgtcgatttc ctaaccttca aagcgactac
     3481 ataactatgt cccctggtat cggcttggat gttaaaggtg acgggctggg acagcaatat
     3541 cgtactcctg aagaagtgat tgtaaactgc ggtagcgata tcatcattgt tggtcgtgga
     3601 gtctatggag ctggtcgtaa tcctgttgtc gaagccaaga gatatagaga agctggttgg
     3661 aaggcatatc agcaaagact ttctcagcat taaaaaaaga ctaatgtaaa atttttttgg
     3721 ttggttattg aaaaagtcga tgccttgttt gcgtttgttt tcctaggcgt tttatgtcag
     3781 aaggcattta gaattagtat acaagtactc tttggtaaaa ttttatgtag cgactaaaat
     3841 attaactatt atagataaac accttgggaa taaaaagtaa tttgctatag taatttatta
     3901 aacatgctcc tacaacatta ccacaatctt ttctcttgga ttgacattga ataagaaaag
     3961 agtgaatttt tttagacttg taatgataac tatgtacaaa gccaatgaaa gatgtatgta
     4021 gatgaatgta aaataccatg tagacaaaca agataaaact tggttataaa cattggtgtt
     4081 ggaacagaat aaattagatg tcaaaaagtt tcgtcaatat cacaagcttc cctgaattcg
     4141 catctagact gatgagacgt ggtagagcca caaacagccg gtacaagcaa cgatctccag
     4201 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
     4261 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat acaacgtgag accgaaagtg
     4321 aaacgtgatt tcatgcgtca ttttgaacat tttgtaaatc ttatttaata atgtgtgcgg
     4381 caattcacat ttaatttatg aatgttttct taacatcgcg gcaactcaag aaacggcagg
     4441 ttcggatctt agctactaga gaaagaggag aaatactaga tgcgtaaagg cgaagagctg
     4501 ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt
     4561 tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc
     4621 tgtactactg gtaaactgcc ggttccttgg ccgactctgg taacgacgct gacttatggt
     4681 gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc
     4741 atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa
     4801 acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc
     4861 attgacttta aagaggacgg caatatcctg ggccataagc tggaatacaa ttttaacagc
     4921 cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt
     4981 cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca
     5041 atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg
     5101 tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg
     5161 ggcatcacgc atggtatgga tgaactgtac aaatgaccag gcatcaaata aaacgaaagg
     5221 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     5281 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggtct cagctggaaa
     5341 tctgctcgtc agtggtgctc acactgacga atcatgtaca gatcataccg atgactgcct
     5401 ggcgactcac aactaagcaa gacagccgga accagcgccg gcgaacacca ctgcatatat
     5461 ggcatatcac aacagtccac gtctcaagca gttacagaga tgttacgaac cactagtgca
     5521 ctgcagtaca gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
     5581 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
     5641 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
     5701 ccgcacggcg cgaagcaaaa attacggctc ctcgctccag acctgcgagc agggaaacgc
     5761 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
     5821 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
     5881 cagttctcac atcacatccg aacataaaca aaaatgggta aggaaaagac tcacgtttcg
     5941 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     6001 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     6061 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     6121 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     6181 gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa
     6241 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     6301 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctggctcag
     6361 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     6421 ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat
     6481 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     6541 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     6601 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     6661 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     6721 taaagtaact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     6781 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     6841 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     6901 tatgtgaatg ctggtcgcta tactggagtt tattattcaa aactagtata ctttttctcg
     6961 gagtataata cacaatatcg gtgcaaatag gttttaaaat tgctccaatc acatgttctt
     7021 aagaaaaaaa cgtcaaaaga aatctaagta ttcccaaaat aattcctcct gacatgagaa
     7081 gaataaaaaa cactaagatg cattgtcagt atgaaacaca agccaaagtt gctaacaata
     7141 ggatcaattc ttacttactg gcaatgttac gaggaatctt ggtaggagga tgtttggcaa
     7201 cataacgttc ttttgaattt gtttttacac gacgagtaat gttcttctga aactgtgtgt
     7261 ttgcacgacg ctgagaggga gtttgaaccg tctaaaagac atagttagga tccatgatgc
     7321 acagatttct aaagataaaa ctacttacag gcatgttggt ttattaaaaa agaaaaacag
     7381 gaatgataag agggaaagaa tacgtgtaca gagca
//
