LOCUS       sequence-213440-        3322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MTS2 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3322)
  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 3322)
  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-MTS2 (12)
FEATURES             Location/Qualifiers
     source          1..3322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..84
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     76..93
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     77..93
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     100..119
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        100..118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      149..158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      153..162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(563..581)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(564..581)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(599..615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(599..615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(612..634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    623..639
                     /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(647..653,654..671,672..677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    692..713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(830..847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1001..1589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1081..1100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1760..2551,2552..2620))
                     /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     2383..2402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2621..2725)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2803..3258)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2940..2961)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3152..3171
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
       61 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
      121 gaattgggcc cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgattggg
      181 cgatgagtgg tctcgccatg gcttctcgga ggcttctcgc ctctctcctc cgtcaatcgg
      241 ctcaacgtgg cggcggtcta atttcccgat cgttaggaaa ctccatccct aaatccgctt
      301 cacgcgcctc ttcacgcgca tcccctaagg gattcctctt aaaccgcgcc gtacagtacg
      361 ctacctccgc agcggcaccg gcatctcagc catcaacacc accaaagtcc ggcagtgaac
      421 cgtccggaaa aattaccgat gagttcaccg gcgctggttc aatgcgagac cactcatcgc
      481 ccaatcacta gtgaattcgc ggccgcctgc aggtcgacca tatgggagag ctcccaacgc
      541 gttggatgca tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt
      601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
      721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
      781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
      841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1441 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1501 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1861 agggcttacc atctggcccc agtgctgcaa tgataccgcg cgacccacgc tcaccggctc
     1921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt
     2761 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     2821 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2881 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2941 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3001 ccgtctatca gggcgctggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3061 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3121 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3181 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3241 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3301 ggtgcgggcc tcttcgctat ta
//
