LOCUS       sequence-386111-        3730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  promoter of the gene alcohol dehydrogenase 2 from R. toruloides fro 
            the P2 position (F/G).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3730)
  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 3730)
  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: (F) pADH2 (G)_P2 (SBE113)
FEATURES             Location/Qualifiers
     source          1..3730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(77..94)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(248..836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(328..347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1007..1798,1799..1867))
                     /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     1630..1649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1868..1972)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2040..2058
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2364..2383
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2364..2382
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(3621..3627,3628..3645,3646..3651))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3666..3687
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
       61 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      121 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      181 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      241 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      301 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      361 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      421 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      481 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      541 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      601 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      661 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      721 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      781 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      961 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1021 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1081 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1141 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1201 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1261 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1321 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1381 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1441 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1501 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1561 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1621 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1681 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1741 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1801 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1861 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1921 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1981 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     2041 ataggcgtat cacgaggccg cccctgcagc cgaattatat tatttttgcc aaataatttt
     2101 taacaaaagc tctgaagtct tcttcattta aattcttaga tgatacttca tctggaaaat
     2161 tgtcccaatt agtagcatca cgctgtgagt aagttctaaa ccattttttt attgttgtat
     2221 tatctctaat cttactactc gatgagtttt cggtattatc tctattttta acttggagca
     2281 ggttccattc attgtttttt tcatcatagt gaataaaatc aactgcttta acacttgtgc
     2341 ctgaacacca tatccatccg gcgtaatacg actcactata gggagagcgg ccgccagatc
     2401 ttccggatgg ctcgagtttt tcagcaagat atgcggtctc attgttgtga ctgtcggaga
     2461 cgtggcagct gtcagctaga cgggaaatga cggtgcgaga gtgaacagcc gggaatggga
     2521 acgcacgaag agagagggct gcgaaggctg gaccggtatg ggctgagtgc gagtgctgtg
     2581 cgagggtgtg tagcagtgtg tgcaagtgaa gagcaagttg taagagagag agcgagcgca
     2641 aggttgcggg acggtcgatg ggtggggaag gcgaagagcg aggaaggcca ctcctgctcg
     2701 ggacgactcg caagtcggct cgctctcgca cgagacacgc ttaattgacg cccacagcca
     2761 ctcgagcacg ggacccagcc accccgaagc aaagcccacc cttcgacgtc tccacccatg
     2821 aattaacgac tgcgggcgag gacggaagac gagtaaagtg tggtggagag cggcggggat
     2881 cagcctagct gccattccca gtgtacacga ctgcaacgac gtcgtcgtag gctctgaccg
     2941 tagtctccga ggtcctactt gatcagtaca ccgcccgcaa caagctcgag ctgttcccac
     3001 accgcgctca cctgctggac tgcctgctac tgcttgctca gcgcacacac accctcaaag
     3061 aacaccgtcg caaaccgcag caccccgccg gcgcgactga tggcatcacg aggaggacga
     3121 ggaggagggc gaggagggag gggaggaggg gcgtcgggga agcctcagcc gaagctgaga
     3181 ccatgcatct ttctagaaga tctcctacaa tattctcagc tgccatggaa aatcgatgtt
     3241 cttcttttat tctctcaaga ttttcaggct gtatattaaa acttatatta agaactatgc
     3301 taaccacctc atcaggaacc gttgtaggtg gcgtgggttt tcttggcaat cgactctcat
     3361 gaaaactacg agctaaatat tcaatatgtt cctcttgacc aactttattc tgcatttttt
     3421 ttgaacgagg tttagagcaa gcttcaggaa actgagacag gaattttatt aaaaatttaa
     3481 attttgaaga aagttcaggg ttaatagcat ccattttttg ctttgcaagt tcctcagcat
     3541 tcttaacaaa agacgtctct tttgacatgt ttaaagttta aacctcctgt gtgaaattat
     3601 tatccgctca taattccaca cattatacga gccggaagca taaagtgtaa agcctggggt
     3661 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgccaat tgctttccag
     3721 tcgggaaacc
//
