LOCUS       sequence-213445-        3464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TDH2t in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3464)
  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 3464)
  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: "BMC 
            Biotechnol."; date: "2017-11-13"; volume: "17(1):80. doi"; pages: " 
            10.1186/s12896-017-0393-y"
COMMENT     Sequence Label: pUPD-TDH2t (17-21)
FEATURES             Location/Qualifiers
     source          1..3464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    127..148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(163..168,169..186,187..193)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    201..217
                     /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     206..228
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     225..241
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     225..241
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        259..277
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     259..276
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      820..829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      824..833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(863..882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(864..882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(889..906)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(889..905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(898..920)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1046..1501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1133..1152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1343..1364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1579..1683
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1684..1752,1753..2544)
                     /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     complement(1902..1921)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2715..3303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3204..3223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3457..3464,1..10)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
       61 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      121 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      181 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      241 ccatgattac gccaagctat ttaggtgaca ctatagaata ctcaagctat gcatccaacg
      301 cgttgggagc tctcccatat ggtcgacctg caggcggccg cgaattcact agtgattggg
      361 cgatgagtgg tctcggcttt aaatttaact ccttaagtta ctttaatgat ttagttttta
      421 ttattaataa ttcatgctca tgacatctca tatacacgtt tataaaactt aaatagattg
      481 aaaatgtatt aaagattcct cagggattcg atttttttgg aagtttttgt ttttttttcc
      541 ttgagatgct gtagtatttg ggaacaatta tacaatcgaa agatatatgc ttacattcga
      601 ccgttttagc cgtgatcatt atcctatagt aacataacct gaagcataac tgacactact
      661 atcatcaata cttgtcacat gagaactctg tgaataatta ggccactgaa atttgatgcc
      721 tgaaggaccg gcatcacgga ttttcgataa agcacttagt atcacactaa ttggcttttc
      781 gccgctcgag accactcatc gcccaatcga attcccgcgg ccgccatggc ggccgggagc
      841 atgcgacgtc gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
      901 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
      961 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1021 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1081 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1141 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1201 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1261 tagggtgatg gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg
     1321 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1381 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1441 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1501 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc
     1561 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1621 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1681 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1741 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1801 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1861 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1921 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1981 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2041 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2101 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2161 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2221 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2281 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2341 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2401 tcgcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2461 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2521 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2581 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2821 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3421 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtc
//
