LOCUS       sequence-199819-        3736 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pTDH3 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3736)
  AUTHORS   Perez-Gonzalez A, Kniewel R, Veldhuizen M, Verma HK, 
            Navarro-Rodriguez M, Rubio LM, Caro E
  TITLE     Adaptation of the GoldenBraid modular cloning system and creation of
            a toolkit for the expression of heterologous proteins in yeast 
            mitochondria.
  JOURNAL   BMC Biotechnol. 2017 Nov 13;17(1):80. doi: 
            10.1186/s12896-017-0393-y.
  PUBMED    29132331
REFERENCE   2  (bases 1 to 3736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol."; date: "2017-11-13"; volume: "17(1):80. doi"; pages: " 
            10.1186/s12896-017-0393-y"
COMMENT     Sequence Label: pUPD-pTDH3 (01-11)
FEATURES             Location/Qualifiers
     source          1..3736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      73..82
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        132..798
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     668..691
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     promoter        complement(901..919)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(902..919)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(937..953)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(937..953)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(950..972)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    961..977
                     /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)."
     promoter        complement(join(985..991,992..1009,1010..1015))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1030..1051
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1168..1185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1339..1927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1419..1438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2098..2889,2890..2958))
                     /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     2721..2740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2959..3063)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3141..3596)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3278..3299)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3490..3509
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3722..3736,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3736,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggcccgacgt
       61 cgcatgctcc cggccgccat ggcggccgcg ggaattcgat tgggcgatga gtggtctcgg
      121 gagcgagttt atcattatca atactgccat ttcaaagaat acgtaaataa ttaatagtag
      181 tgattttcct aactttattt agtcaaaaaa ttagcctttt aattctgctg taacccgtac
      241 atgcccaaaa tagggggcgg gttacacaga atatataaca tcgtaggtgt ctgggtgaac
      301 agtttattcc tggcatccac taaatataat ggagcccgct ttttaagctg gcatccagaa
      361 aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc aaccatcagt tcataggtcc
      421 attctcttag cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct
      481 caatggagtg atgcaacctg cctggagtaa atgatgacac aaggcaattg acccacgcat
      541 gtatctatct cattttctta caccttctat taccttctgc tctctctgat ttggaaaaag
      601 ctgaaaaaaa aggttgaaac cagttccctg aaattattcc cctacttgac taataagtat
      661 ataaagacgg taggtattga ttgtaattct gtaaatctat ttcttaaact tcttaaattc
      721 tacttttata gttagtcttt tttttagttt taaaacacca agaacttagt ttcgaataaa
      781 cacacataaa caaacaaacc atcgagacca ctcatcgccc aatcactagt gaattcgcgg
      841 ccgcctgcag gtcgaccata tgggagagct cccaacgcgt tggatgcata gcttgagtat
      901 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      961 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1021 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1081 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1141 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1201 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1741 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2221 tgctgcaatg ataccgcgcg acccacgctc accggctcca gatttatcag caataaacca
     2281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2461 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2701 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3001 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3061 gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa
     3121 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3181 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3241 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3301 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgctggccc
     3361 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3421 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3481 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3541 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     3601 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3661 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3721 tcccagtcac gacgtt
//
