LOCUS       Exported                4218 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Part of GoldenGate Yarrowia lipolytica toolkit for assembling an 
            expression plasmid for Y. lipolytic yeast. Coding sequence of 
            fluorescent protein in position 1 (G1_RedStarII).
ACCESSION   .
VERSION     .
KEYWORDS    GGE077
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4218)
  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 4218)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4218
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        143..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(1106..1125)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1107..1125)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1132..1149)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1132..1148)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1141..1163)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1286..1588
                     /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     1467..1486
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1937..2731
                     /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(1991..2010)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2601..2620
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2931..2940
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2937..3311
                     /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      3449..4037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3938..3957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4191..4208
                     /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 tcttagatta caagaacaag
      361 tggtgtctac cttcagcacg ttcgtattgt tcgacgatag tgtagtcttc gttgtgagag
      421 atgatgtcca acttagagtc aacgtagtaa tagcctggca attggacttg cttcttggcc
      481 ttgtagatag acttgaattc gaccaagtag tgaccaccgt ccttcaactt caaggccttg
      541 tggatttcac ccttcaagac accgtctctt gggtacaaac gttcggtaga tggttcccaa
      601 cccatagtct tcttttgcat gactggacca tcagatggga agttgacacc aatgagcttg
      661 accttgtaga tgaaacaacc gtcttgcaag gaggagtctt gagtaacagt aacaacgcca
      721 ccgtcttcga agttcatgat cctttcccac ttgaaacctt ctgggaagga caacttcttg
      781 tagtctggaa tgtcagctgg gtgcttgacg tagaccttag aaccgtattg gaattgtgga
      841 gacaagatgt cccaagcgaa tggcagtgga ccacccttag taaccttcaa cttagctgtt
      901 tgagtacctt catatggtct accctcacct tcaccttcga tttcgaattc gtggccgtta
      961 acagtacctt ccattttaac cttgaatctc atgaattcag tgatgacatc ttcagaagaa
     1021 gcactcatta gagaccaagg gcgaattctg cagatatcca tcacactggc ggccgctcga
     1081 gcatgcatct agagggccca attcgcccta tagtgagtcg tattacaatt cactggccgt
     1141 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     1201 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1261 acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa gagagagccg
     1321 ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc gacggatggt
     1381 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     1441 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     1501 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     1561 cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa aaaggatctt
     1621 cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     1681 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     1741 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     1801 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     1861 ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg
     1921 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     1981 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2041 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2101 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2161 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2221 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2281 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2341 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     2401 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2461 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     2521 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     2581 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     2641 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     2701 cttctatcgc cttcttgacg agttcttctg aattattaac gcttacaatt tcctgatgcg
     2761 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc acttttcggg
     2821 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2881 tcatgagaca ataaccctga taaatgcttc aataatagca cgtgaggagg gccaccatgg
     2941 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     3001 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     3061 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     3121 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     3181 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     3241 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     3301 agcaggactg acacgtgcta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4201 tgagcgagga agcggaag
//
