LOCUS       sequence-259155-        3900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of GoldenGate Yarrowia lipolytica toolkit for assembling an 
            expression plasmid for Y. lipolytic yeast. Promoter for gene 
            expression in position1 (P1_Tef).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3900)
  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 3900)
  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: GGE085
FEATURES             Location/Qualifiers
     source          1..3900
                     /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(788..807)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(789..807)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(814..831)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(814..830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(823..845)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             968..1270
                     /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     1149..1168
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1619..2413
                     /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             1619..2413
                     /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(1673..1692)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2283..2302
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2613..2622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2619..2993
                     /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      3131..3719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3620..3639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3873..3890
                     /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 gcccttggtc tctacggggg ttggcggcgt
      361 atttgtgtcc caaaaaacag ccccaattgc cccaattgac cccaaattga cccagtagcg
      421 ggcccaaccc cggcgagagc ccccttcacc ccacatatca aacctccccc ggttcccaca
      481 cttgccgtta agggcgtagg gtactgcagt ctggaatcta cgcttgttca gactttgtac
      541 tagtttcttt gtctggccat ccgggtaacc catgccggac gcaaaataga ctactgaaaa
      601 tttttttgct ttgtggttgg gactttagcc aagggtataa aagaccaccg tccccgaatt
      661 acctttcctc ttcttttctc tctctccttg tcaactcaca cccgaagaat gagagaccaa
      721 gggcgaattc tgcagatatc catcacactg gcggccgctc gagcatgcat ctagagggcc
      781 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
      841 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      901 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctata
      961 cgtacggcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     1021 tgtacagagt gatattattg acacgccggg gcgacggatg gtgatccccc tggccagtgc
     1081 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     1141 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     1201 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     1261 gggaatataa atgtcaggca tgagattatc aaaaaggatc ttcacctaga tccttttcac
     1321 gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat
     1381 ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg
     1441 gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc
     1501 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag
     1561 gatctgatgg cgcaggggat caagctctga tcaagagaca ggatgaggat cgtttcgcat
     1621 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     1681 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     1741 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     1801 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     1861 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     1921 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     1981 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2041 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2101 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     2161 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     2221 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     2281 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     2341 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     2401 cgagttcttc tgaattatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     2461 tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2581 gataaatgct tcaataatag cacgtgagga gggccaccat ggccaagttg accagtgccg
     2641 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     2701 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     2761 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     2821 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     2881 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     2941 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
     3001 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3061 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3241 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3781 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3841 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
