LOCUS       sequence-386063-        4714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  promoter of the gene alcohol dehydrogenase 2 from R. toruloides for 
            the position P1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4714)
  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 4714)
  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: (R)pADH2(D)_P1(SBE214)
FEATURES             Location/Qualifiers
     source          1..4714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1086..1105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1087..1105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1112..1129)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1112..1128)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1121..1143)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1266..1568
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1447..1466
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1917..2711
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1917..2711
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1971..1990)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2581..2600
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(2961..3752,3753..3821))
                     /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     3584..3603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3945..4533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4434..4453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4687..4704
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc ccttagagca
      301 ggtctccgtg tcggctgagg cttccccgac gcccctcctc ccctccctcc ccgccctcct
      361 cctcgtcctc ctcgtgatgc catcagtcgc gccggcgggg tgctgcggtt tgcgacggtg
      421 ttctttgagg gtgtgtgtgc gctgagcaag cagtagcagg cagtccagca ggtgagcgcg
      481 gtgtgggaac agctcgagct tgttgcgggc ggtgtactga tcaagtagga cctcggagac
      541 tacggtcaga gcctacgacg acgtcgttgc agtcgtgtac actgggaatg gcagctaggc
      601 tgatccccgc cgctctccac cacactttac tcgtcttccg tcctcgcccg cagtcgttaa
      661 ttcatgggtg gggacgtcga agggtgggct ttgcttcggg gtggctgggt cccgtgctcg
      721 agtggctgtg ggcgtcaatt aagcgtgtct cgtgcgagag cgagccgact tgcgagtcgt
      781 cccgagcagg agtggccttc ctcgctcttc gccttcccca cccatcgacc gtcccgcaac
      841 cttgcgctcg ctctctctct tacagcttgc tcttcacttg cacacactgc tacacaccct
      901 cgcacagcac tcgcactcag cccataccgg tccagccttc gcagccctct ctcttcgtgc
      961 gttcccattc ccggctgttc actctcgcac cgtcatttcc cgtctagctg acagctgcca
     1021 cgtctccgac agtcacaaat gtgagaccgc ataagggcga attcgcggcc gctaaattca
     1081 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     1141 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     1201 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg
     1261 tacggcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     1321 tacagagtga tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac
     1381 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     1441 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     1501 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     1561 gaatataaat gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt
     1621 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     1681 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     1741 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     1801 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     1861 tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga
     1921 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     1981 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2041 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2101 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2161 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2221 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2281 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2341 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2401 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     2461 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2521 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2581 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2641 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2701 agttcttctg aattattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     2761 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagatt atcaaaaagg
     2881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2941 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3001 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3061 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3121 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3181 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3241 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3301 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3361 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3421 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3481 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3541 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3601 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3661 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3721 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3781 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3841 tattgaagca tttatcaggg ttattgtctc atgaccaaaa tcccttaacg tgagttttcg
     3901 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3961 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4021 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4081 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4141 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4201 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4261 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4321 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4381 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4441 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4501 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4561 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4621 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4681 gagcgcagcg agtcagtgag cgaggaagcg gaag
//
