LOCUS       sequence-213444-        3461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADH2t in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3461)
  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 3461)
  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-ADH2t (17-21)
FEATURES             Location/Qualifiers
     source          1..3461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        652..670
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     652..669
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      776..1075
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     regulatory      1210..1219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1214..1223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1253..1272)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1254..1272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1279..1296)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1279..1295)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1288..1310)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1436..1891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1523..1542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1733..1754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1969..2073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2074..2142,2143..2934)
                     /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(2292..2311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3105..3461,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tatttaggtg
      661 acactataga atactcaagc tatgcatcca acgcgttggg agctctccca tatggtcgac
      721 ctgcaggcgg ccgcgaattc actagtgatt gggcgatgag tggtctcggc tttaagcgga
      781 tctcttatgt ctttacgatt tatagttttc attatcaagt atgcctatat tagtatatag
      841 catctttaga tgacagtgtt cgaagtttca cgaataaaag ataatattct actttttgct
      901 cccaccgcgt ttgctagcac gagtgaacac catccctcgc ctgtgagttg tacccattcc
      961 tctaaactgt agacatggta gcttcagcag tgttcgttat gtacggcatc ctccaacaaa
     1021 cagtcggtta tagtttgtcc tgctcctctg aatcgtatcc ctcgatattt ctcattttcc
     1081 ttcgcatgcc agcattgaaa tgatcgaagt tcaatgatga aacggtaatt cttctgtcat
     1141 ttactcatct catctcatca agttatataa ttcgctcgag accactcatc gcccaatcga
     1201 attcccgcgg ccgccatggc ggccgggagc atgcgacgtc gggcccaatt cgccctatag
     1261 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     1321 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     1381 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     1441 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1501 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1561 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1621 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccagcg
     1681 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1741 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1801 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1861 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     1921 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2281 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2581 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2761 gctgataaat ctggagccgg tgagcgtggg tcgcgcggta tcattgcagc actggggcca
     2821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3241 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa c
//
