LOCUS       sequence-258851-        3237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of GoldenGate Yarrowia lipolytica toolkit for assembling an 
            expression plasmid for Y. lipolytic yeast. Destination vector for 
            the insertion of the assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3237)
  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 3237)
  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: GGE0029
FEATURES             Location/Qualifiers
     source          1..3237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    94..115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(130..135,136..153,154..160)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    168..184
                     /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)."
     RBS             202..213
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             220..897
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      942..1013
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1029..1056
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1093..1136
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1140..1158)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1226..1330
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1331..1399,1400..2191)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1549..1568)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2362..2950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2851..2870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3104..3121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3145..3202)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
ORIGIN
        1 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
       61 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      121 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      181 acaatttcac acatactaga gaaagaggag aaatactaga tggcttcctc cgaagacgtt
      241 atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg tcacgagttc
      301 gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc taaactgaaa
      361 gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca gttccagtac
      421 ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa actgtccttc
      481 ccggaaggtt tcaaatggga acgtgttatg aacttcgaag acggtggtgt tgttaccgtt
      541 acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact gcgtggtacc
      601 aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga agcttccacc
      661 gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg tctgaaactg
      721 aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa aaaaccggtt
      781 cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca caacgaagac
      841 tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg tgcttaataa
      901 cgctgatagt gctagtgtag atcgctacta gagccaggca tcaaataaaa cgaaaggctc
      961 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     1021 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taactagagg tctcaaggct
     1081 cttcgaattc cagaaatcat ccttagcgaa agctaaggat tttttttatc tgaaattctg
     1141 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1201 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1261 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1321 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1381 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1441 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1501 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1561 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1621 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1681 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1741 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1801 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1861 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1921 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1981 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2041 gcgtgggtcg cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2101 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2161 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2221 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2281 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2341 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2401 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2461 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2521 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2581 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2641 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2701 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2761 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2821 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2881 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2941 cctgtggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3001 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3061 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3121 ggaagcctgc ataacgcgaa gtaatctttt cggttttaaa gaaaaagggc agggtggtga
     3181 caccttgccc ttttttgccg gactgcagga agacgcagga gaccactaga caatacg
//
