LOCUS       sequence-428699-        7356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty integration vector for genomic integration of 1 
            transcriptional unit 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 7356)
  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 7356)
  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: pPOM041_P15678_SingleGeneBackbone
FEATURES             Location/Qualifiers
     source          1..7356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(107..898,899..967))
                     /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     730..749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(968..1072)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1175..1763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1664..1683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2840..3634
                     /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             4424..4435
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4442..5155
                     /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      5167..5238
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5254..5281
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     gene            5511..6866
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     CDS             5855..6664
                     /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(5922..5941)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6669..6866
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 ccgagcggcc gcgattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
       61 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      241 taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1081 cccgaaaagt gccacctgtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1141 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1201 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1261 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1321 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1381 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1501 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1561 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1621 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1681 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1741 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1801 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1861 gtgcggccgc caatatgctt acataatgag tgagtgtcat tattgaggaa tgccatgtca
     1921 gatttgacac aactcagaac ttattaactt tttatgattg gaacttttgt tttgcccaag
     1981 gaatttttaa tggaagaagt tgttttataa atgatattga atttatatcc actcttttta
     2041 gcgcatttgc atctctactt taggttcttc tttgactaga ttctaggatg cgtattggat
     2101 atactggtct ttgaatcatt gtcgaaaaaa aaattcatgg ttgtacccat gagcaaactg
     2161 cttttgtggt tatccatgtg cttcttttgt tcagcatcta tcttcttaat cgcatggaag
     2221 atttgattta aaccaaaaga aatattgtta aactttgaca gaagctttaa attttgattc
     2281 catacattaa atggataaaa ctcctgaagc ttagctacaa atcccactgg ctatatgtat
     2341 gcatttgtgt taaaaaagtt tgtatagatt atttaatcta ctcagcattc tttctctaaa
     2401 taggaatttg ttacttaatg gagaaaaaaa tgtttcgatt tacctagtgt atttgtttgt
     2461 atactcacgt ttaatttcaa acatccattc tatcttgtgt aatttttggc atggtgaaaa
     2521 agataatcag ccttataatc tttacaaaag taagaaattc tgtaaataag ccttaatgcc
     2581 cttgctttaa attaaaatgg ttctttttca tgataatgtt tgcactttgt gaatatattt
     2641 tagatagttc tgtgaggtat aattaagatg ttttagagac ttatacaatt ttgtctttat
     2701 aaattcttaa ttgattttac catcccagtt taactatgct tcgtcggcat ctctgcacat
     2761 gtcgtgtttt cttaccgtat tgtcctacca agaacctctt ttttgcttgg atcgaaatta
     2821 aaggtttaaa agcaaagtta tggatgctag agtatttcaa agctattcag ctagagctga
     2881 ggggatgaaa aatcccattg ccaaggaatt gttggctttg atggaagaaa agcaaagcaa
     2941 cttgtcagtc gcggtcgatt tgacgaagaa atccgaaatc ttagaattgg tagataaaat
     3001 tggaccctat gtctgtgtta tcaagacaca tattgacgtt gtcgaggatt tcgaccagga
     3061 tatggtagaa aaactggtgg ccttaggtaa aaagcatcgt tttcttatct ttgaggatcg
     3121 caaattcgca gacattggaa ataccgtcaa gctacaatat gcatctggtg tgtacaaaat
     3181 tgcttcttgg gctcatatca caaattgcca tacagtgcca ggcgagggta ttatacaagg
     3241 cctcaaagaa gttggtttac ctttgggacg tggactcttg cttttggctg aaatgtcttc
     3301 caaaggctct ttggctactg gttcctacac agagaaaacc ttagaatggt ttgagaagca
     3361 taccgatttt tgctttggct ttatagctgg tcgtcgattt cctaaccttc aaagcgacta
     3421 cataactatg tcccctggta tcggcttgga tgttaaaggt gacgggctgg gacagcaata
     3481 tcgtactcct gaagaagtga ttgtaaactg cggtagcgat atcatcattg ttggtcgtgg
     3541 agtctatgga gctggtcgta atcctgttgt cgaagccaag agatatagag aagctggttg
     3601 gaaggcatat cagcaaagac tttctcagca ttaaaaaaag actaatgtaa aatttttttg
     3661 gttggttatt gaaaaagtcg atgccttgtt tgcgtttgtt ttcctaggcg ttttatgtca
     3721 gaaggcattt agaattagta tacaagtact ctttggtaaa attttatgta gcgactaaaa
     3781 tattaactat tatagataaa caccttggga ataaaaagta atttgctata gtaatttatt
     3841 aaacatgctc ctacaacatt accacaatct tttctcttgg attgacattg aataagaaaa
     3901 gagtgaattt ttttagactt gtaatgataa ctatgtacaa agccaatgaa agatgtatgt
     3961 agatgaatgt aaaataccat gtagacaaac aagataaaac ttggttataa acattggtgt
     4021 tggaacagaa taaattagat gtcaaaaagt ttcgtcaata tcacaagctt ccctgaattc
     4081 gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag gaccatctga
     4141 atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc acagtacaga
     4201 cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc gataattgca
     4261 gacgaacgtg agaccgaaag tgaaacgtga tttcatgcgt cattttgaac attttgtaaa
     4321 tcttatttaa taatgtgtgc ggcaattcac atttaattta tgaatgtttt cttaacatcg
     4381 cggcaactca agaaacggca ggttcggatc ttagctacta gagaaagagg agaaatacta
     4441 gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
     4501 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
     4561 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct
     4621 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
     4681 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
     4741 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt
     4801 aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa
     4861 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
     4921 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
     4981 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
     5041 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
     5101 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc
     5161 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     5221 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     5281 gtttataggt ctcagctgag catgagacgg aaatctgctc gtcagtggtg ctcacactga
     5341 cgaatcatgt acagatcata ccgatgactg cctggcgact cacaactaag caagacagcc
     5401 ggaaccagcg ccggcgaaca ccactgcata tatggcatat cacaacagtc caactagtgc
     5461 actgcagtac agtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg
     5521 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
     5581 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
     5641 gccgcacggc gcgaagcaaa aattacggct cctcgctcca gacctgcgag cagggaaacg
     5701 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     5761 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
     5821 acagttctca catcacatcc gaacataaac aaaaatgggt aaggaaaaga ctcacgtttc
     5881 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     5941 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     6001 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     6061 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     6121 tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga
     6181 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     6241 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctggctca
     6301 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     6361 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
     6421 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     6481 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     6541 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     6601 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     6661 ctaaagtaac tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt
     6721 tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc
     6781 tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc
     6841 gtatgtgaat gctggtcgct atactggagt ttattattca aaactagtat actttttctc
     6901 ggagtataat acacaatatc ggtgcaaata ggttttaaaa ttgctccaat cacatgttct
     6961 taagaaaaaa acgtcaaaag aaatctaagt attcccaaaa taattcctcc tgacatgaga
     7021 agaataaaaa acactaagat gcattgtcag tatgaaacac aagccaaagt tgctaacaat
     7081 aggatcaatt cttacttact ggcaatgtta cgaggaatct tggtaggagg atgtttggca
     7141 acataacgtt cttttgaatt tgtttttaca cgacgagtaa tgttcttctg aaactgtgtg
     7201 tttgcacgac gctgagaggg agtttgaacc gtctaaaaga catagttagg atccatgatg
     7261 cacagatttc taaagataaa actacttaca ggcatgttgg tttattaaaa aagaaaaaca
     7321 ggaatgataa gagggaaaga atacgtgtac agagca
//
