LOCUS       sequence-386109-        5579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rhodotorula toruloides gene phytoene dehydrogenase (crtI, 
            RHTO_04602) for G2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5579)
  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 5579)
  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: (G) crtI (H) G2 (SBE116)
FEATURES             Location/Qualifiers
     source          1..5579
                     /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(3411..3417,3418..3435,3436..3441))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3456..3477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3597..3614)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3768..4356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3848..3867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4527..5318,5319..5387))
                     /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     5150..5169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5388..5492)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5560..5578
                     /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 caatcctcaa ccgcgcaggt acatcaccaa caaagcaatc
      421 aacccaaaca acaccgagag gagcgtaaac cctcccgctg catcgcgggt cgtcctgtgc
      481 gtcgcgagtt ccgaggagga gacgttccag cgcgagggga tgggcatccc gaggtcgttg
      541 aggatttggg ttgcgacgag gcgggcgccg gcgaggacga tggggacgct gcgaggacaa
      601 tacgaattag caaccattcc ttgaactcga cgcatgcggt acgctggaga ggcagcgaca
      661 agcggggcga gcgaagacca ccacaactca cccagttccc gggtgcgccg acgcgccgac
      721 aaagtaagcg ttcttgacac tcgggtggcg ggtcttgggg cggaaagaga ggacgttgct
      781 gcgaagaagc gaaaggttgg gaaggtcagc gcaagcttca agagcaagca ggacaagaca
      841 ggctggcgga actcacaaga agtcgtgact gagtccgagg atgctgccgc ggtgcaaatt
      901 gaacttctcg ccccaagtga tgggcgtgtt gatcgtctcg tgctcgatgc agctccggag
      961 gtcctcgatg tcgaggcggc gctcaatctc gccgatgatc ttgttgcgcg tctcttcgac
     1021 gaccttgtcc cagtcggaag aggagggggg ggcggcggag atgtgcccga cgggaacgag
     1081 gacgatgacg gcgtccttgt cggcgggtgc ggcgctgcga gcgggtgcgg agtcagcgat
     1141 ggccggctgc tgacgacgtg gaagggacat acgacgggtc gtgacggctg ggaacgttga
     1201 cgtagaagga aggctcatgc gggacgcggt gttcgcggaa gatcgagtcg aaggacctgg
     1261 acgcgagagg cttcgtcagc gaccgcttcg agagcggtga gtagcgcagg tgtgctcact
     1321 ctcggtactc ctctgcgagg aagatggtat gcgagtccag ctggggtatc ttgctgtagc
     1381 ggccagaggg gagtaaggat ccgccattcg cgatcgagga atagagtgtc ttcacgcacc
     1441 ggttcatggg ccagtagaac gagatggacg agcagctgac gggccgctcc tcgagcctct
     1501 tcgagtagct cgtctcctcg tacaggtgcg ccatactcca caccaaatcc gcattgacga
     1561 ggacgatgtc ggcggtcaac ttctcgccgc tctcgagctc gacaccctta gccgtgccgt
     1621 tctcgaccgt gtcgcgcttg acgggactgt tgaagcggaa cttggcgccg ctcttttcgg
     1681 cgagctgttg gagggtttgc gggatgcggc cgaagccgcc gagagggtag aggatgcctt
     1741 cgcagtgctc ggtgtactgg aggaggttgt aggcgccgag agcgtcgaag ggggacatgc
     1801 cctgcgagat tggaggtgcg ggtgagcgga ggaggaactt ggccggcgat taaagccaga
     1861 gctggcgctc gaatcgcgac gctctccgac tcacgagata catcgaggtg aaagtgaacg
     1921 ctctccgcat gcggtccgtc ttgaagtact tggtcgcgcg cgagtactga gtcgaggcga
     1981 cgctcatcag caatcctgcg ttcccagcga gttgtatcag cactgatgag gtgacgtacg
     2041 acagagacaa agggatggag cttgcggagc tggatgatga gagacgggcg gaccatcgag
     2101 aggagcgagg tgaagttgcg gtgcaggacg tggaccatcg acagctcgta gtgcgcgtgt
     2161 ccctccttca agaagccaag gaatctgcat acaacatgag cgtcagtaac tggaaaggaa
     2221 gcgggaagag agcacaagcg aaagggcatc tccgctcgtc gtctcgctcc gaggttccaa
     2281 gccactcacc cttcaaagcc cttctctccc tcccaccgct cgacctgctt cttcaccctt
     2341 gtcaagtcgc tgctcatctc gacgacctcc ttgtcgggga agacgatcca gtagtttggc
     2401 tcgcacttga cgagcctgat gccctcgttc tcgaggctcg tcccaagatc gttaaagatc
     2461 tcgtcaaaca gtcgcgggat gaggaacagg ctcgggccct ggtcgaagcg gaaggacttg
     2521 gtcggatcgg tgaagaggga gcatcgtccg ccgacaaagt cgttcttctc gaggactatc
     2581 gacgaggttg aactcgtcag tgcctccaac caggttctac gagtgctccc acgtcccccg
     2641 atcgaggcag gacgacgcag gagacagagt gtgcgcacct gtcacgtcga acccggattg
     2701 ggcgaggcga gcggcggagg cagtccctcc gacgcccgct ccgacgatga gcaccgaggg
     2761 ctttcccttg ccgtgtccgt tcgcagctgc catccttgct gtgctaatga gttggtggca
     2821 ggacaagagc aaccccgtca agcatcacac cgccccgcct ggcatcggcg atcgagcgga
     2881 tgagttcccg gaagcgatca tctccacctt gcgttgctcg gcatcgcatc gaagctcctc
     2941 ttcacgtgca agcgggcgca tttgttgaga ccatgcatct ttctagaaga tctcctacaa
     3001 tattctcagc tgccatggaa aatcgatgtt cttcttttat tctctcaaga ttttcaggct
     3061 gtatattaaa acttatatta agaactatgc taaccacctc atcaggaacc gttgtaggtg
     3121 gcgtgggttt tcttggcaat cgactctcat gaaaactacg agctaaatat tcaatatgtt
     3181 cctcttgacc aactttattc tgcatttttt ttgaacgagg tttagagcaa gcttcaggaa
     3241 actgagacag gaattttatt aaaaatttaa attttgaaga aagttcaggg ttaatagcat
     3301 ccattttttg ctttgcaagt tcctcagcat tcttaacaaa agacgtctct tttgacatgt
     3361 ttaaagttta aacctcctgt gtgaaattat tatccgctca taattccaca cattatacga
     3421 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3481 gcgttgcgct cactgccaat tgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3541 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3601 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3661 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3721 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3781 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3841 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3901 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3961 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4021 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4081 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4141 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4201 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4261 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4321 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4381 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4441 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4501 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4561 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4621 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4681 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4741 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4801 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4861 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4921 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4981 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5041 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5101 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5161 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5221 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5281 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5341 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5401 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5461 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5521 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggcc
//
