LOCUS       sequence-263227-        3192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of GoldenGate Yarrowia lipolytica toolkit for assembling an 
            expression plasmid for Y. lipolytic yeast. Sequence for homologous 
            integration into Y. lipolytica genome. Linearization of cassette by 
            NotI restriction enzyme after the assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3192)
  AUTHORS   Larroude M, Park YK, Soudier P, Kubiak M, Nicaud JM, Rossignol T
  TITLE     A modular Golden Gate toolkit for Yarrowia lipolytica synthetic 
            biology.
  JOURNAL   Microb Biotechnol. 2019 May 31. doi: 10.1111/1751-7915.13427.
  PUBMED    31148366
REFERENCE   2  (bases 1 to 3192)
  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: "Microb 
            Biotechnol. 2019 May 31. doi: 10.1111/1751-7915.13427."
COMMENT     Sequence Label: GGE207
FEATURES             Location/Qualifiers
     source          1..3192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(79..125)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(126..142)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(126..142)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(252..271)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             263..1078
                     /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             263..1078
                     /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     complement(336..355)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1214..1802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1703..1722
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1955..1977
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1974..1990
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1974..1990
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3126..3143)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3126..3142)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3135..3157)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gagacttcct tggccgtacg atgggttttg tttcaggtat cccaacctgg acgtctcgta
       61 taatgcgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
      121 atttggatcc actagttcta gagcggcggg ggaaagccac gttgtgtctc aaaatctctg
      181 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
      241 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc
      301 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
      361 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
      421 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
      481 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
      541 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
      601 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
      661 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
      721 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
      781 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
      841 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
      901 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
      961 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1021 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1081 agaatttgtt aattggttgt aacactggca gagcggacga tttgaagccc cctacccatt
     1141 tacgctgact tgacgggacg gcggctttgt tgaatagagc gtcagacccc gtagaaaaga
     1201 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1261 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1321 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1381 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1441 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1501 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1561 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1621 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1681 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1741 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1801 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1861 tgttctttcc agcgacatat gccatacgcc gaactgaact ccacctaatg tcttggagat
     1921 tgtcggatag cgcggaatag gattgtgccc tctgagcgga taacaatttc acacaggaaa
     1981 cagctatgac catgacacat gcatccacca tcgcagactt atcatcactg agcctccacc
     2041 tagcctgggg tctctcgcat gcttgaaggc atatgtgact gttatttgct ggttcatgag
     2101 atactgtttg tccattgatt ccaatgcacg gtctgaatgt tcaaaatcag taaaagacac
     2161 aaaaccgtac ccttttgaag aaccatctgt gtcacgagat attcgagggg gtttggcaaa
     2221 tggtccaaac acagagaacg tttgctgtag tgtagactca tctacgagag gatccaaact
     2281 tcccacgaaa agtgttgctc caatctcaaa cccgttttgc ttgtcggatg cagatttgct
     2341 gactcgaatg gattttccga acagccagat ttggttcata atagacgctg cgtaatttgc
     2401 atcagcctct gttttgtact ccacgaagcc atagccttgg tgtgtctggc tgactctgtc
     2461 tttaggaaga ttgatgtgca cgatagggcc cgcttgaagc ataagttcat acagaatcgc
     2521 ttctgtgacg ttctcgtcaa ggttgcctat ataaagagta gcctcagggt tctggtccgc
     2581 ttcttgttga ttcactctgt tcatgaaagc tgtcgatttg gcgcgatgtg tgagaattca
     2641 acacgattat gcacgatcaa tttaggttgt tagtaataaa aaagtcccac cacgaagtct
     2701 cttgatcgca acttgtttta ttttattttt tcatccataa aaatttttat acctgtaata
     2761 acattatatt gggggagggg gcgagatcca atatatatat tttataatca cataatacag
     2821 tacaagtatt cgtacagtac ttgaacaaca ctttgttgaa ctattaacac atcagaccct
     2881 ctactgatct cacacttccg gtacagtatg tactgtactt gcagtacaca gcgcgataca
     2941 ttactctacg ttttttttta tggtggtggt cctaactgaa actataacag tattaactaa
     3001 tcttaactca tacatttctt aaaatttact taactcgtat cacttggtca gataatagca
     3061 ctcagagacc cccaggtcag gcggaatggc acttccttat tccagatgcg tgcggattat
     3121 gcatgactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgc ttgtggtgga
     3181 tcacagttat tg
//
