LOCUS       sequence-416652-        3790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  promoter of the gene glyceraldehyde-3-phosphate dehydrogenase from 
            R. toruloides for a P2 position (overhangs F/G).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3790)
  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 3790)
  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)pGPD(G)_P2 (SBE110)
FEATURES             Location/Qualifiers
     source          1..3790
                     /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(1622..1628,1629..1646,1647..1652))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1667..1688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1808..1825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1979..2567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2059..2078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2738..3529,3530..3598))
                     /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     3361..3380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3599..3703)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3771..3789
                     /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 tgcatggtct cagctttgtg actgatctgg tgttgttctg agcgggtaga
      421 gcgagagcgg gtgatgggaa agagtgcaaa gggacgcgaa cttgtcgatg cggcttcctt
      481 cgctcacgaa atcagccctc gcgcgtttcg tcccccggct ggacccgacc tgcgctcgag
      541 gcgggcgtgt gacgacttcc tgcgctgcag attcagacgg atcatgctct cgagggattg
      601 tttccctcct tcaccgcttc gctcgccctc tctgcgacgt cggctgagat cgataagttg
      661 gtgatgtgac gtggccatct ccgaggttcc tttccgaccc ttacaagccg agattcgcgc
      721 agcttcgatt gattgtattg ctcgaggtaa ccgagtatga gggagtcgtc tctcgtcgac
      781 cccaagcctg cgcatactgc tgcccgcttc tggtgcctgc tccaataaat tactgtactt
      841 ggcgccgcga tttctggact atctcgcttg ccttatgctc ccgcggcgcc gctctgcgcc
      901 gttcctccca cgacttccat gtcctcacag cttgtttcgt cgtccgaagc gtcctctacg
      961 tcttgcaaac gtcaaaaaac gacgttggca tcccggcggc acaaggcttg ctccggtagg
     1021 cgacctacaa ggcgagccag cagatttgct tgtccgcaga gatttcttcc ttccatcgtg
     1081 ccgtctaaag gacgaaacga ggtcacggag gagggaccta gagagcaagg ttcattccaa
     1141 tgcttgcgga gcgaggagaa cgtcgaactc caacaatgag accgcatatc tttctagaag
     1201 atctcctaca atattctcag ctgccatgga aaatcgatgt tcttctttta ttctctcaag
     1261 attttcaggc tgtatattaa aacttatatt aagaactatg ctaaccacct catcaggaac
     1321 cgttgtaggt ggcgtgggtt ttcttggcaa tcgactctca tgaaaactac gagctaaata
     1381 ttcaatatgt tcctcttgac caactttatt ctgcattttt tttgaacgag gtttagagca
     1441 agcttcagga aactgagaca ggaattttat taaaaattta aattttgaag aaagttcagg
     1501 gttaatagca tccatttttt gctttgcaag ttcctcagca ttcttaacaa aagacgtctc
     1561 ttttgacatg tttaaagttt aaacctcctg tgtgaaatta ttatccgctc ataattccac
     1621 acattatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1681 tcacattaat tgcgttgcgc tcactgccaa ttgctttcca gtcgggaaac ctgtcgtgcc
     1741 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1801 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2401 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3601 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3661 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3721 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3781 tcacgaggcc
//
