LOCUS       sequence-262907-        3946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of GoldenGate Yarrowia lipolytica toolkit for assembling an 
            expression plasmid for Y. lipolytic yeast. Inducible promoter for 
            gene expression in position 2 (P2_EYK300-3AB).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3946)
  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 3946)
  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: GGE108
FEATURES             Location/Qualifiers
     source          1..3946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    174..190
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     179..201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     198..214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     198..214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        232..250
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     232..249
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(834..853)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(835..853)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(860..877)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(860..876)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(869..891)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1014..1316
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1195..1214
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1665..2459
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1665..2459
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1719..1738)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2329..2348
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2659..2668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2665..3039
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3177..3765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3666..3685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3919..3936
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttatgct tccggctcgt atgttgtgtg gaattgtgag
      181 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tatttaggtg
      241 acactataga atactcaagc tatgcatcaa gcttggtacc gagctcggat ccactagtaa
      301 cggccgccag tgtgctggaa ttcgcccttg cggtctcttt gtggatccag tagatgtgta
      361 agtgtgtaga agtgtcgtgt gggccaattg tggggaaagg agcttcttat agaagctgat
      421 ctgcccctct ttctgatcca cgttgacccc cacgctgttt tcagctttgc aaaagtttgg
      481 gaccgcggta cgtggtggcc ttgcctcgaa ataatagtct ttgtccgcac tagtcaggtc
      541 atcggattat gcaaggctgt ggagcttaat gcggcggctt tcccttttgg gattccgctt
      601 cccgctgtgg agcttaatgc ggcggctttc ccttttggga ttccgcttcc cgctgtggag
      661 cttaatgcgg cggctttccc ttttgggatt ccgcttcccc agattgaaac gtacagtata
      721 gagaaaagta gatgcatcga tacgcgtatc gaagcagaga ccgcaagggc gaattctgca
      781 gatatccatc acactggcgg ccgctcgagc atgcatctag agggcccaat tcgccctata
      841 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
      901 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
      961 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctatacgta cggcagttta
     1021 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     1081 ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     1141 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     1201 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     1261 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     1321 caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag aaagccagtc
     1381 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     1441 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     1501 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     1561 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     1621 ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     1681 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     1741 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     1801 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     1861 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     1921 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     1981 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     2041 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2101 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     2161 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     2221 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     2281 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     2341 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     2401 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa
     2461 ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2521 cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2581 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2641 taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc ggtgctcacc
     2701 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     2761 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     2821 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     2881 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     2941 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     3001 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctaaa acttcatttt
     3061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3301 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3841 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3901 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
