LOCUS       sequence-259323-        4343 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 position 2 (P2_PGM).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4343)
  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 4343)
  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: GGE004
FEATURES             Location/Qualifiers
     source          1..4343
                     /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(1231..1250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1232..1250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1257..1274)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1257..1273)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1266..1288)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1411..1713
                     /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     1592..1611
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2062..2856
                     /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             2062..2856
                     /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(2116..2135)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2726..2745
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3056..3065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3062..3436
                     /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      3574..4162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4063..4082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4316..4333
                     /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 tctgctttac caaccacaga
      361 ttacgaccca ttcgcagtca cagttcacta gggtttgggt tgcatccgtt gagagtggtt
      421 tgtttttaac cttctccatg tgctcactca ggttttgggt tcagatcaaa tcaaggcgtg
      481 aaccactgtt tgaggacaaa tgtgacacaa ccaaccagtg tcaggggcaa gtccgtgaca
      541 aaggggaaga tacaatgcaa ttactgacag ttacggactg cctcgatgcc ctaaccttgc
      601 cccaaaataa gacaactgtc ctcgtttaag cgcaacccta ttcagcgtca cgtcataata
      661 gcgtttggat agcactagtc tatgaggagc gttttatgtt gcggtgaggg cgattggtgc
      721 tcatatgggt tcaattgagg tggtggaacg agcttagtct tcaattgagg tgcgagcgac
      781 acaattgggt gtcacgtggc ctaattgacc tcggatcgtg gagtccccag ttatacagca
      841 accacgaggt gcatgagtag gagacgtcac cagacaatag ggtttttttg gactggagag
      901 ggtagggcaa aagcgctcaa cgggctgttt ggggagctat gggggaggaa ttggcgatat
      961 ttgtgaggtt gacggctccg atttgcgtgt tttgtcgctt ctgcatctcc ccatacccat
     1021 atcttccctc cccacctctt tccacgataa ttttacggat cagcaataag gttccttctc
     1081 ctagtttcca cgtccatata tatctatgct gcgtcgtcct tttcgtgaca tcaccaaaac
     1141 acatacaaaa acaaagagac caagggcgaa ttctgcagat atccatcaca ctggcggccg
     1201 ctcgagcatg catctagagg gcccaattcg ccctatagtg agtcgtatta caattcactg
     1261 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1321 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1381 tcccaacagt tgcgcagcct atacgtacgg cagtttaagg tttacaccta taaaagagag
     1441 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc ggggcgacgg
     1501 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac
     1561 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     1621 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     1681 aacgccatta acctgatgtt ctggggaata taaatgtcag gcatgagatt atcaaaaagg
     1741 atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     1801 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag
     1861 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc
     1921 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     1981 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag
     2041 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2101 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2161 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2221 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     2281 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2341 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2401 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2461 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     2521 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     2581 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     2641 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     2701 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     2761 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     2821 atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta caatttcctg
     2881 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt
     2941 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3001 tccgctcatg agacaataac cctgataaat gcttcaataa tagcacgtga ggagggccac
     3061 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     3121 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     3181 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     3241 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     3301 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     3361 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     3421 cgaggagcag gactgacacg tgctaaaact tcatttttaa tttaaaagga tctaggtgaa
     3481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3721 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4201 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4261 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4321 gtcagtgagc gaggaagcgg aag
//
