LOCUS       sequence-110256-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pre-Assembled Ura3 integration vector to be used in the Dueber YTK 
            system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5283)
  AUTHORS   Lee ME, DeLoache WC, Cervantes B, Dueber JE
  TITLE     A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
  JOURNAL   ACS Synth Biol. 2015 Apr 14.
  PUBMED    25871405
REFERENCE   2  (bases 1 to 5283)
  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. 2015 Apr 14."
COMMENT     Sequence Label: pYTK096
FEATURES             Location/Qualifiers
     source          1..5283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             865..876
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             883..1596
                     /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      1608..1679
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1695..1722
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2111..2914
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(3556..4371)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3556..4371)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4279..4298
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4363..4382
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4596..5184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5085..5104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcca atgctaaatt cgagtgaaac acaggaagat cagaaaatcc tcatttcatc
       61 catattaaca ataatttcaa atgtttattt gcattatttg aaactaggca agacaagcaa
      121 cgaaacgttt ttgaaaattt tgagtatttt caataaattt gtagaggact cagatattga
      181 aaaaaagcta cagcaattaa tacttgataa gaagagtatt gagaagggca acggttcatc
      241 atctcatgga tctgcacatg aacaaacacc agagtcaaac gacgttgaaa ttgaggctac
      301 tgcgccaatt gatgacaata cagacgatga taacaaaccg aagttatctg atgtagaaaa
      361 ggattaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt ctatatatgt
      421 taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc atcaaagaag
      481 gttaatgtgg ctgtggtttc agggtccata aagcccacat ggataacatt acccctgaat
      541 tcgcatctag actgatgaga cgtggtagag ccacaaacag ccggtacaag caacgatctc
      601 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
      661 cccacagtac agacactgcg acaacgtggc aattcgtcgc aatacaacgt gagaccgaaa
      721 gtgaaacgtg atttcatgcg tcattttgaa cattttgtaa atcttattta ataatgtgtg
      781 cggcaattca catttaattt atgaatgttt tcttaacatc gcggcaactc aagaaacggc
      841 aggttcggat cttagctact agagaaagag gagaaatact agatgcgtaa aggcgaagag
      901 ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag
      961 ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc
     1021 atctgtacta ctggtaaact gccggttcct tggccgactc tggtaacgac gctgacttat
     1081 ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc
     1141 gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac
     1201 aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa
     1261 ggcattgact ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac
     1321 agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa
     1381 attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact
     1441 ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt
     1501 ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca
     1561 gcgggcatca cgcatggtat ggatgaactg tacaaatgac caggcatcaa ataaaacgaa
     1621 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     1681 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagg tctcagctgg
     1741 aaatctgctc gtcagtggtg ctcacactga cgaatcatgt acagatcata ccgatgactg
     1801 cctggcgact cacaactaag caagacagcc ggaaccagcg ccggcgaaca ccactgcata
     1861 tatggcatat cacaacagtc cacgtctcaa gcagttacag agatgttacg aaccactagt
     1921 gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga
     1981 agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga
     2041 aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg
     2101 aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt
     2161 tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga
     2221 tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt
     2281 actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct
     2341 aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat
     2401 tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga
     2461 cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc
     2521 agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg
     2581 ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga
     2641 ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg
     2701 gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta
     2761 tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact
     2821 atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga
     2881 agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg
     2941 tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtagag
     3001 cacttgaatc cactgccccg ggaatctcgg tcgtaatgat ttctataatg acgaaaaaaa
     3061 aaaaattgga aagaaaaagc ttcatggcct ttataaaaag gaactatcca atacctcgcc
     3121 agaaccaagt aacagtattt tacggggcac aaatcaagaa caataagaca ggactgtaaa
     3181 gatggacgca ttgaactcca aagaacaaca agagttccaa aaagtagtgg aacaaaagca
     3241 aatgaaggat ttcatgcgtt tgtactctaa tctggtagaa agatgtttca cagactgtgt
     3301 caatgacttc acaacatcaa agctaaccaa taaggaacaa acatgcatca tgaagtgctc
     3361 agaaaagttc ttgaagcata gcgaacgtgt agggcagcgt ttccaagaac aaaacgctgc
     3421 cttgggacaa ggcttgggcc gataaggtgt actggcgtat atatatctaa ttatgtatct
     3481 ctggtgtagc ccatttttag catgtaaata taaagaccga gcggccgcgt gttacaacca
     3541 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     3601 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     3661 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     3721 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     3781 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     3841 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     3901 attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt
     3961 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4021 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     4081 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     4141 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     4201 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     4261 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     4321 ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct
     4381 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     4441 tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt
     4501 tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga
     4561 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4621 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4681 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4741 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4801 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4861 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4921 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4981 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5041 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5101 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5161 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5221 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5281 cgt
//
