LOCUS       sequence-259427-        5783 bp    DNA     circular SYN 15-APR-2025
DEFINITION  loxP-KIURA3-loxP preassembled entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5783)
  AUTHORS   An-Adirekkun JM, Stewart CJ, Geller SH, Patel MT, Melendez J, Oakes 
            BL, Noyes MB, McClean MN
  TITLE     A yeast optogenetic toolkit (yOTK) for gene expression control in 
            Saccharomyces cerevisiae.
  JOURNAL   Biotechnol Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234.
  PUBMED    31788779
REFERENCE   2  (bases 1 to 5783)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234."
COMMENT     Sequence Label: pMM0617
FEATURES             Location/Qualifiers
     source          1..5783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             875..886
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             893..1606
                     /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      1618..1689
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1705..1732
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    1964..1997
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3449..3482
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4066..4881)
                     /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(4066..4881)
                     /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     4789..4808
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4873..4892
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5106..5694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5595..5614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt gcggccgcca atgctaaatt cgagtgaaac acaggaagat cagaaaatcc
       61 tcatttcatc catattaaca ataatttcaa atgtttattt gcattatttg aaactaggca
      121 agacaagcaa cgaaacgttt ttgaaaattt tgagtatttt caataaattt gtagaggact
      181 cagatattga aaaaaagcta cagcaattaa tacttgataa gaagagtatt gagaagggca
      241 acggttcatc atctcatgga tctgcacatg aacaaacacc agagtcaaac gacgttgaaa
      301 ttgaggctac tgcgccaatt gatgacaata cagacgatga taacaaaccg aagttatctg
      361 atgtagaaaa ggattaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt
      421 ctatatatgt taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc
      481 atcaaagaag gttaatgtgg ctgtggtttc agggtccata aagcccacat ggataacatt
      541 acccctgaat tcgcatctag actgatgaga cgtggtagag ccacaaacag ccggtacaag
      601 caacgatctc caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca
      661 cagacattaa cccacagtac agacactgcg acaacgtggc aattcgtcgc aatacaacgt
      721 gagaccgaaa gtgaaacgtg atttcatgcg tcattttgaa cattttgtaa atcttattta
      781 ataatgtgtg cggcaattca catttaattt atgaatgttt tcttaacatc gcggcaactc
      841 aagaaacggc aggttcggat cttagctact agagaaagag gagaaatact agatgcgtaa
      901 aggcgaagag ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa
      961 cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac
     1021 gctgaagttc atctgtacta ctggtaaact gccggttcct tggccgactc tggtaacgac
     1081 gctgacttat ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt
     1141 cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga
     1201 cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat
     1261 tgagctgaaa ggcattgact ttaaagagga cggcaatatc ctgggccata agctggaata
     1321 caattttaac agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc
     1381 gaattttaaa attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca
     1441 gcaaaacact ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac
     1501 gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt
     1561 cgtaaccgca gcgggcatca cgcatggtat ggatgaactg tacaaatgac caggcatcaa
     1621 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1681 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagg
     1741 tctcagctgg aaatctgctc gtcagtggtg ctcacactga cgaatcatgt acagatcata
     1801 ccgatgactg cctggcgact cacaactaag caagacagcc ggaaccagcg ccggcgaaca
     1861 ccactgcata tatggcatat cacaacagtc cacgtctcaa gcagttacag agatgttacg
     1921 aaccactagt gcactgcagt acagcaggtc gacaaccctt aatataactt cgtataatgt
     1981 atgctatacg aagttattag gtctagagat cccaatacaa cagatcacgt gatctttttg
     2041 taagatgaag ttgaagtagt gttgcaccgt gccaatgcag gtggctatta gattaaatat
     2101 gtgatttgtt ctattaagtt tcctgtataa ttaatgggga gcgctgattc tcttttggta
     2161 cgcttcccat ccagcatttc tgtatctttc accttcaacc ttaggatctc tacccttggc
     2221 gaaaagtcct ctgccaacaa tgatgatatc tgatccacca cttacaactt cgtcgacggt
     2281 tctgtactgc tgacccaatg catcgccttt gtcgtctaaa cctacacctg gggtcatgat
     2341 tagccaatca aacccttctt ctcttcctcc catatcgttc tgagcaatga acccaataac
     2401 gaaatcttta tcactctttg caatatcaac ggtaccctta gtatattcac cgtgtgctag
     2461 agaacccttg gaagacaatt cagcaagcat caataatccc cttggttctt tggtgacctc
     2521 ttgcgcacct tgtttcaagc cagcaacaat accagcacca gtaaccccgt gggcgttggt
     2581 gatatcagac cattctgcga tacggtaaac gcccgatgta tattgtaatt tgactgtgtt
     2641 accgatatcg gcgaattttc tgtcctcaaa tatcaagaac ttgtatttct ctgccaatgc
     2701 tttcaatgga acgacagtac cctcataact gaaatcatcc aagatatcaa cgtgtgtttt
     2761 caaaaggcaa atgtatggac ccaacgtttc aacaagtttc aatagctcat cagtcgaacg
     2821 aacgtcaaga gaagcacaca aattggtctt cttttcatcc attaaacgta aaagtttcga
     2881 tgcaaccgga cttgcatgag tctcagctct actggtatat gattttgtgg acatggtgca
     2941 actaattgac gggagtgtat tgacgctggc gtactggctt tcacaaaatg gcccaatcac
     3001 aaccacatct tagatagttg aaatgacttt agataacatc aattgagatg agcttaatca
     3061 tgtcaaagct aaaagtgtca ccatgaacga caattcttaa gcaaatcacg tgatatagat
     3121 ccacgaataa ccaccatttg atgctcgagg caagtaatgt gtgtaaaaaa atgcgttacc
     3181 accatccaat gcagaccgat cttctaccca gaatcacata tatttatgta ccgagtacct
     3241 tttttctatc ttccaattgc ttctcccata tgattgtctc cgtaagctcg aaatttctaa
     3301 gttggatttt aatcttcacg caggatgaca gttcgatgag cttctgagga gtgtttagaa
     3361 cataatcatt tatccatggt ctatctcttc ttgtcgcttt ttctcctcga tagaacctaa
     3421 ataaaacgag ctctcgagaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     3481 attaggtgat atcagatcca ctgagtagag cacttgaatc cactgccccg ggaatctcgg
     3541 tcgtaatgat ttctataatg acgaaaaaaa aaaaattgga aagaaaaagc ttcatggcct
     3601 ttataaaaag gaactatcca atacctcgcc agaaccaagt aacagtattt tacggggcac
     3661 aaatcaagaa caataagaca ggactgtaaa gatggacgca ttgaactcca aagaacaaca
     3721 agagttccaa aaagtagtgg aacaaaagca aatgaaggat ttcatgcgtt tgtactctaa
     3781 tctggtagaa agatgtttca cagactgtgt caatgacttc acaacatcaa agctaaccaa
     3841 taaggaacaa acatgcatca tgaagtgctc agaaaagttc ttgaagcata gcgaacgtgt
     3901 agggcagcgt ttccaagaac aaaacgctgc cttgggacaa ggcttgggcc gataaggtgt
     3961 actggcgtat atatatctaa ttatgtatct ctggtgtagc ccatttttag catgtaaata
     4021 taaagaccga gcggccgcgt gttacaacca attaaccaat tctgattaga aaaactcatc
     4081 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     4141 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     4201 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     4261 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     4321 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     4381 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg
     4441 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     4501 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     4561 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     4621 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     4681 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     4741 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     4801 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg agcaagacgt
     4861 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     4921 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     4981 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagt catgaccaaa
     5041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5221 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5761 cctgcgttat cccctgattc tgt
//
