LOCUS       sequence-369785-        2921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.4 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2921)
  AUTHORS   Shaw WM, Khalil AS, Ellis T
  TITLE     A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423.
  PUBMED    37930278
REFERENCE   2  (bases 1 to 2921)
  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 6. doi: 10.1021/acssynbio.3c00423."
COMMENT     Sequence Label: pMYT088
FEATURES             Location/Qualifiers
     source          1..2921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(869..1684)
                     /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(869..1684)
                     /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     1592..1611
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1676..1695
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1909..2497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2398..2417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcagtacgtt gaaaatacat tcagtcacac ccgcagcaca gcaagtcgtt tcatctaagc
       61 tattttgatc gttagaatat ccacacctca caagtatcga caatttagat gtgtattggg
      121 ttttgtctac gcaaatacca ttagtttcac acaagatgcc atcagcgcta ctgtaaatag
      181 attgttcctc tgggcttgga gtgagagtga cagaattcgc attctagacg agcatgacac
      241 tatctgcgcg gactagttag cctgcagcat gtacaatgtc ttccgatgaa gacatgagtc
      301 gacacggacc tgagttagtt tatcgtgcga atggaactag aaaaccaata atcattgaat
      361 aatatatgtg acagtgacag gccaccagaa taaagctaat atctggaggg gtatttcttc
      421 ttgagtgaag aagaaatatg ctacttccaa attttaggac gacaaatcct cataatatga
      481 attaggcaaa ttacagaatg aagggggttg agaagtccca cagcacatcg ctccttcgct
      541 aagagctcca ctgctaaaac agaaaaaaaa agtattaatc aattgtacta ccatgttctt
      601 tgttactaaa cgcgcctatt tttttcgaga gcctttacga gagtcaatca agcattctcc
      661 cgcatacttc gctaagaaaa aagtgatctt taattgggcc cttcatctat catttttgct
      721 taagtggcgt aacttatttt atcatttcct ctccctgcca tcctctttat ttttgttctg
      781 tatcacagca gttaattcag tttccattca aagtatgcca cggataactt gatgcggccg
      841 cgtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
      901 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
      961 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     1021 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     1081 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     1141 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     1201 caaccaaacc gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt
     1261 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     1321 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     1381 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     1441 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     1501 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     1561 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     1621 cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc
     1681 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     1741 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     1801 taaatcgaac ttttgctgag ttgaaggatc agtcatgacc aaaatccctt aacgtgagtt
     1861 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1921 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1981 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2041 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2101 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2161 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2221 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2281 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2341 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2401 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2461 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2521 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2581 ttctgtggat aaccgtgcgg ccgcaataca ctaaattgtc catgttttaa ctaccatttc
     2641 cttctaattt ttttcctttc ttagctgatt taattcattt atttgctgtc gtttgcattt
     2701 actgactaaa aaaagtgtaa aatatgggaa gcgcaaattg aaaagtcggg ttgcgaaatt
     2761 acctgccaaa aatccacctc gttaatatct aacaagattc aacgcctgtt gacaaaagaa
     2821 actcgtcagt ctctcagttt acttctatgt ttaacatatt tgctatgatc cactccagat
     2881 tctgcccagc cctcaacttc acgcccacgt tgagagtaca t
//
