LOCUS       sequence-386107-        3730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  promoter of the gene alcohol dehydrogenase 2 from R. toruloides for 
            the P3 position (I/J).
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: (I) pADH2 (J) P3 (SBE114)
FEATURES             Location/Qualifiers
     source          1..3730
                     /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(1562..1568,1569..1586,1587..1592))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1607..1628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1748..1765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1919..2507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1999..2018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2678..3469,3470..3538))
                     /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     3301..3320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3539..3643)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3711..3729
                     /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 cagtggtgtg actgtcggag acgtggcagc tgtcagctag
      421 acgggaaatg acggtgcgag agtgaacagc cgggaatggg aacgcacgaa gagagagggc
      481 tgcgaaggct ggaccggtat gggctgagtg cgagtgctgt gcgagggtgt gtagcagtgt
      541 gtgcaagtga agagcaagtt gtaagagaga gagcgagcgc aaggttgcgg gacggtcgat
      601 gggtggggaa ggcgaagagc gaggaaggcc actcctgctc gggacgactc gcaagtcggc
      661 tcgctctcgc acgagacacg cttaattgac gcccacagcc actcgagcac gggacccagc
      721 caccccgaag caaagcccac ccttcgacgt ctccacccat gaattaacga ctgcgggcga
      781 ggacggaaga cgagtaaagt gtggtggaga gcggcgggga tcagcctagc tgccattccc
      841 agtgtacacg actgcaacga cgtcgtcgta ggctctgacc gtagtctccg aggtcctact
      901 tgatcagtac accgcccgca acaagctcga gctgttccca caccgcgctc acctgctgga
      961 ctgcctgcta ctgcttgctc agcgcacaca caccctcaaa gaacaccgtc gcaaaccgca
     1021 gcaccccgcc ggcgcgactg atggcatcac gaggaggacg aggaggaggg cgaggaggga
     1081 ggggaggagg ggcgtcgggg aagcctcagc cgtgactgag accatgcatc tttctagaag
     1141 atctcctaca atattctcag ctgccatgga aaatcgatgt tcttctttta ttctctcaag
     1201 attttcaggc tgtatattaa aacttatatt aagaactatg ctaaccacct catcaggaac
     1261 cgttgtaggt ggcgtgggtt ttcttggcaa tcgactctca tgaaaactac gagctaaata
     1321 ttcaatatgt tcctcttgac caactttatt ctgcattttt tttgaacgag gtttagagca
     1381 agcttcagga aactgagaca ggaattttat taaaaattta aattttgaag aaagttcagg
     1441 gttaatagca tccatttttt gctttgcaag ttcctcagca ttcttaacaa aagacgtctc
     1501 ttttgacatg tttaaagttt aaacctcctg tgtgaaatta ttatccgctc ataattccac
     1561 acattatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1621 tcacattaat tgcgttgcgc tcactgccaa ttgctttcca gtcgggaaac ctgtcgtgcc
     1681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1741 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1861 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2521 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2581 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2641 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2701 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2761 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2821 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2881 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2941 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3001 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3061 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3121 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3181 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3241 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3301 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3361 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3421 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3481 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3541 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3601 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3661 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3721 tcacgaggcc
//
