LOCUS       sequence-263181-        4265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of GoldenGate Yarrowia lipolytica toolkit for assembling an 
            expression plasmid for Y. lipolytic yeast. Coding sequence of 
            fluorescent protein in position 3 (G3_mTurquoise).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4265)
  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 4265)
  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: GGE261
FEATURES             Location/Qualifiers
     source          1..4265
                     /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."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1153..1172)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1154..1172)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1179..1196)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1179..1195)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1188..1210)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1333..1635
                     /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     1514..1533
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1984..2778
                     /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             1984..2778
                     /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(2038..2057)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2648..2667
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2978..2987
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2984..3358
                     /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      3496..4084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3985..4004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4238..4255
                     /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 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttgggg tctctccaca tggtttcaaa
      361 gggagaggaa ctgttcacag gcgtcgtccc gatacttgtg gaacttgacg gtgacgtaaa
      421 cggacataag ttttctgtct ctggtgaggg tgaaggtgac gccacctacg gaaagctgac
      481 tctgaaattc atttgcacca ccggaaagct tccagttcct tggcctaccc tggtcactac
      541 cctgtcttgg ggtgtgcaat gtttcgctcg ataccccgat cacatgaagc agcacgactt
      601 tttcaagtct gccatgcccg agggatacgt tcaggagcga actatcttct tcaaggacga
      661 tggtaactac aagactcgag ctgaggttaa gttcgagggt gacactctgg tcaaccgaat
      721 tgaacttaag ggtattgact tcaaggagga tggcaacatt ctgggtcaca agctggagta
      781 taactacttt agtgacaatg tctacatcac cgcggataag cagaagaacg gaatcaaagc
      841 caactttaag attagacata atattgagga tgggggagtg cagctggccg atcactacca
      901 gcagaacact cccatcggtg acggccccgt cctgctgcct gacaaccact acctctcgac
      961 ccagagcaag ttgtccaagg accctaacga gaagcgtgac cacatggtgc tcctggagtt
     1021 tgtgacagct gctggtatca ccctcggcat ggacgagctc tacaagtaag tatagagacc
     1081 gggaagggcg aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga
     1141 gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     1201 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     1261 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     1321 ctatacgtac ggcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     1381 gtggatgtac agagtgatat tattgacacg ccggggcgac ggatggtgat ccccctggcc
     1441 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     1501 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     1561 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     1621 ttctggggaa tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1681 ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     1741 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     1801 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     1861 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     1921 ccaaggatct gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt
     1981 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2041 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2101 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2161 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2221 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2281 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2341 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2401 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2461 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     2521 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2581 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     2641 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     2701 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     2761 cttgacgagt tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt
     2821 acgcatctgt gcggtatttc acaccgcatc aggtggcact tttcggggaa atgtgcgcgg
     2881 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2941 accctgataa atgcttcaat aatagcacgt gaggagggcc accatggcca agttgaccag
     3001 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     3061 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     3121 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     3181 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     3241 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     3301 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     3361 cgtgctaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3601 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4201 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4261 ggaag
//
