LOCUS       sequence-386110-        4509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rhodotorula toruloides gene geranylgeranyl diphosphate synthase 
            (crtE, RHTO_02504) for G1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4509)
  AUTHORS   Bonturi N, Pinheiro MJ, de Oliveira PM, Rusadze E, Eichinger T, 
            Liudziute G, De Biaggi JS, Brauer A, Remm M, Miranda EA, 
            Ledesma-Amaro R, Lahtvee PJ
  TITLE     Development of a dedicated Golden Gate Assembly Platform (RtGGA) for
            Rhodotorula toruloides.
  JOURNAL   Metab Eng Commun. 2022 May 23;15:e00200. doi: 
            10.1016/j.mec.2022.e00200. eCollection 2022 Dec.
  PUBMED    35662893
REFERENCE   2  (bases 1 to 4509)
  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: "Metab Eng 
            Commun."; date: "2022-05-23"; volume: "15:e00200. doi"; pages: " 
            10.1016/j.mec.2022.e00200. eCollection 2022 Dec"
COMMENT     Sequence Label: (D) crtE (E)_G1 (SBE115)
FEATURES             Location/Qualifiers
     source          1..4509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(2341..2347,2348..2365,2366..2371))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2386..2407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2527..2544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2698..3286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2778..2797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3457..4248,4249..4317))
                     /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     4080..4099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4318..4422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4490..4508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tatgcggtct catagatcag actttgggaa gctcgtgcgc atggacgcca
      421 ttgacgccgt tcgtcaccgc cttgaccggt ttgctcttga caacctcctc cagcttcctc
      481 tccatcgcct cgccctcaac gccagtcgcc ctcgcctgcg gttcctccaa cacaagcttg
      541 tcgagaatcg cctcgacgcc tctgttccct cccagccgcg caacctcatc cctcgcctgc
      601 tgcgtcaact tgcgcaacac ctcgcgcgtg tacgcgaagg agcccgtccg cgtctccatg
      661 tagctgacag cgtactcttt cggaccggga gaagagggcc gctcgcggag gatgttgagg
      721 atttggcggt tcgaggtgtc ggagcggatc gagtggacga tggggaagga gaatttgcct
      781 tcggagaagt cttcgcagaa gcccttgttg tttgcgtact gtaagaatga tgtatgagca
      841 actttgggca gctcggagcg agcggaaaac gcacctcgac cgattgcagg ttcacgtagt
      901 cgtcgcgaat ctggaagatg atgccgatca ggttggcgag cgggacgtaa tctctgggag
      961 cacaatcgtc agtcctaatc tatgcgagcg gaattaagcc agcgcaccgt ggtgctgcag
     1021 gcgaagcggc catcatcaac ttgatcgcga tgcggaacag tcctcccgtc tctgcgatcc
     1081 cgaccgagtc agcgcacgcc gtatagaata gactcgcacg acgcactgtt gttgaccatg
     1141 tcgatgtact ccggttctgt cgggcagatc aagttctcgc gccagaagag gtccatccct
     1201 tggccgcgat gcaggttcag cagctcctct gcggggatcg cgtcagtctc caaggcttcg
     1261 acaaggttga ggcgaaccca ccggtgacca tctcttcgac cttgatgccc ggccgcgggt
     1321 gtatccgctg gagttcttga aacgcgagaa agtagacata gttggccgag ttgatcgtct
     1381 ggggaatccc gtagatcttg tgtgcgactg gaggatggcg cacgggatca gtctctgcga
     1441 cgagaggcgg ctcggagatc ggagactgac caggcatgcc tcgacggagg tgcgagtcgt
     1501 cctcgacgtc gtccatcctg cgggaatcga cgcggatcgt gagcacccgg tattgctgga
     1561 cagcgagagt gtctcgagag cgcacagcag gctggccgtg tgcagcatcc cgacgacttt
     1621 cttgacaatc tcgagatcga cgtctgcgac acccatccac tggttgaacg ccgcgatgag
     1681 cgccgagcgc acttccttgc ccgggatcga ggcgaggtag gtgtacggct ctgtcaggac
     1741 ctgcacacga gcgtgtcagc gctggcttcg acgactcgcg aggatgtggg gaggcgacgg
     1801 cggtgccggg cgggcgcacc tgctcctgcg cgggctgcca cgacggcgtg ccctgcacct
     1861 tgtcgataaa gttgtcgtac cagtccagcg acatttgaga ccatgcatct ttctagaaga
     1921 tctcctacaa tattctcagc tgccatggaa aatcgatgtt cttcttttat tctctcaaga
     1981 ttttcaggct gtatattaaa acttatatta agaactatgc taaccacctc atcaggaacc
     2041 gttgtaggtg gcgtgggttt tcttggcaat cgactctcat gaaaactacg agctaaatat
     2101 tcaatatgtt cctcttgacc aactttattc tgcatttttt ttgaacgagg tttagagcaa
     2161 gcttcaggaa actgagacag gaattttatt aaaaatttaa attttgaaga aagttcaggg
     2221 ttaatagcat ccattttttg ctttgcaagt tcctcagcat tcttaacaaa agacgtctct
     2281 tttgacatgt ttaaagttta aacctcctgt gtgaaattat tatccgctca taattccaca
     2341 cattatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2401 cacattaatt gcgttgcgct cactgccaat tgctttccag tcgggaaacc tgtcgtgcca
     2461 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2521 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2581 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3121 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3541 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3601 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3661 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3721 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3781 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3841 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3901 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3961 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4021 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4081 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4141 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4201 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4261 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4321 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4381 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4441 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4501 cacgaggcc
//
