LOCUS       sequence-386095-        5256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rhodotorula toruloides gene phytoene synthase/lycopene cyclase 
            (crtYB, RHTO_04605) for G3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5256)
  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 5256)
  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: (J) crtYB(K) G3 (SBE117)
FEATURES             Location/Qualifiers
     source          1..5256
                     /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(3088..3094,3095..3112,3113..3118))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3133..3154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3274..3291)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3445..4033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3525..3544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4204..4995,4996..5064))
                     /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     4827..4846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5065..5169)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5237..5255
                     /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 caatactcac agcgcctgcc acgctttgcg cgccctcgca
      421 cccttggaca ccctcgcgcg ctccatcacc ctcccctcgt ccaacgcctt cccgaccgcg
      481 tccccgatgc tgagatagac caaacacgcc gccctgatcc ctcctctcgc ctcttgcggc
      541 aacttctcta tcgcatcctt gctatgcgca gccatctcgt tcgcacgaga gagcaacgcc
      601 cgcctgtcgc tcgtcaactc gggcaccggc gtgtcgagcg cgcgacctgg gaggtagatc
      661 cgcccaatct tcaagtcggc cggtacgtcc cgcgcgatat tgacgagctg gagagcctgt
      721 cccatctccc tcgccgccgc gagaacgtgc gcgttctccg cttctgacag aatcgggtcg
      781 cgcgggactg attgagcggg tgtgcgggag taaggcgtgc agtgcgccca gacgagttgg
      841 acgcagagat cggcgacgga cgaggcgacg ctgttggcgt agacgagcag gtccgagtcg
      901 gtcttgatgg gcaactcggc ggagatgttc gacttcccgt tgccgttcgc gctaccgttc
      961 gtgtgcgcgc ccttcgagtc ggagagagcg agaaaggaga ggtcggtgcg gaagccgtcg
     1021 aggagctcgt cgagcggtgg gcgggcgatc ggcagaagac tgaggaggcg gaaagtgcct
     1081 cgttcgggct cgctcaaacc gggcaaggcg gcctcgatct gcttgttcga gacgcggacg
     1141 gggagggagg tgggcgtgaa ggaggaggga gggtagagga ggtcgaggca gcccgaaatc
     1201 atgtcgatgt tggattcggc cgccgcgacc gtcgaggcgt tgtcgatcag gtcgtcgcaa
     1261 actcggcacc aggcgtagct gcggtattct gaagatcagc tctgatctat gcttcgatac
     1321 gagccgcact cacagggaga gaaggtccag cctcagcctg ccctcgaaca cctgactcgc
     1381 cgtcgagaag ctcttcgaat ggaccgacaa gatgtcgata gccgcctgca agtcctcgat
     1441 gcgttgacgc tgtgcggcgt cgggcgagtt gaggaggatg gggaggaagt cgcgcaaggg
     1501 tgggaagacg gacgaggtgc gggtgtcgta ggagcgcaag tcgtgaactg caaggcagta
     1561 gtcgcaggcg acgaggccga agacgatcat gacgtttgtc aagaggaaga agacggcctc
     1621 tctgcgtgaa cgttgaccgt cagcaagcga ctgggcgatg ataggcagac cgggcgcact
     1681 cgatttcaag gccgcggaaa gccaagccca gctttgtgcc cttctcgatc acccaagttc
     1741 cgcgctgtaa tgctcgagcg tcgcactccc acaagtagag cgtcgggagg aggatgggaa
     1801 gaccgacggc ccagcgaggc atggcgagca agtggttcga ggcaatgaac ctgcaaggga
     1861 caagtcagtc atcactctcg ccgggaactc gcttggacgc accagagcaa cgcgaggaac
     1921 ggcgcaaccc atccaacaat caacgcgagg tacgttccct ctcctccctc ttccaacttg
     1981 gcccacgcga gcgcaaagat gcccaagaac gccgccgtcc cgatgtaccg cgccgctcgt
     2041 ccgtcgctag gtttccgagg gaggaggacg gcgtgcacga cggggcggct gaagagggcg
     2101 tagacggtcg cggtgatgta ggtctggacg aagaagaaga agatctcttc gtagggtatc
     2161 gcgaagaggg tcggtccgac gacggatgac tcggggtatg accagatctg tccggtgcaa
     2221 gcgttagtcg gcgctttcga gggtaggaag caagacttgg cgcaccctgt gccggatgag
     2281 gtacgagtcc cagggaattg tcgcctgcgc gagattcggt cagttcctcg cgtctcctgc
     2341 ttcttgtgag ctgcacgaac ggttactgcg atctgtggga cgggcagagg tcagttggcg
     2401 attggagcat cgagccgacg gaggaagcgg tgtggacgca cggcgacggg gaagagcgtc
     2461 ttgtacacat cgcggcgtgt cctgagcttc ctgaaggtgc tccagaggac tagcgctggc
     2521 gggatagtcc agcggaggtg gctgtcgggc gttgcgagca acacgagggt gagtgcggag
     2581 gatgcgagtg aacgggagcg aggtggacgc acacgagcca gtagtccagt ccgcccatgt
     2641 ggtgagacca tgcatctttc tagaagatct cctacaatat tctcagctgc catggaaaat
     2701 cgatgttctt cttttattct ctcaagattt tcaggctgta tattaaaact tatattaaga
     2761 actatgctaa ccacctcatc aggaaccgtt gtaggtggcg tgggttttct tggcaatcga
     2821 ctctcatgaa aactacgagc taaatattca atatgttcct cttgaccaac tttattctgc
     2881 attttttttg aacgaggttt agagcaagct tcaggaaact gagacaggaa ttttattaaa
     2941 aatttaaatt ttgaagaaag ttcagggtta atagcatcca ttttttgctt tgcaagttcc
     3001 tcagcattct taacaaaaga cgtctctttt gacatgttta aagtttaaac ctcctgtgtg
     3061 aaattattat ccgctcataa ttccacacat tatacgagcc ggaagcataa agtgtaaagc
     3121 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgccaattgc
     3181 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3241 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3301 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3361 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3421 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3481 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3541 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3601 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3661 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3721 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3781 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3841 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3901 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3961 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4021 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4081 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4141 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4201 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4261 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4321 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4381 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4441 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4501 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4561 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4621 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4681 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4741 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4801 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4861 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4921 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4981 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5041 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5101 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5161 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5221 gacattaacc tataaaaata ggcgtatcac gaggcc
//
