LOCUS       sequence-365746-        6173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation of mitochondrial DNA in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6173)
  AUTHORS   Rzepka M, Suhm T, Ott M
  TITLE     Incorporation of reporter genes into mitochondrial DNA in budding 
            yeast.
  JOURNAL   STAR Protoc. 2022 May 25;3(2):101359. doi: 
            10.1016/j.xpro.2022.101359. eCollection 2022 Jun 17.
  PUBMED    35634362
REFERENCE   2  (bases 1 to 6173)
  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: "STAR 
            Protoc."; date: "2022-05-25"; volume: "3(2):101359. doi"; pages: " 
            10.1016/j.xpro.2022.101359. eCollection 2022 Jun 17"
COMMENT     Sequence Label: pPT24-mtnluc
FEATURES             Location/Qualifiers
     source          1..6173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(251..267)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(252..268)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(296..314)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(335..351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(335..351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(348..370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    359..375
                     /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(383..389,390..407,408..413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    428..449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(566..583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(737..1325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(817..836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1496..2287,2288..2356))
                     /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     2119..2138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2357..2461)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2487..2942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2624..2645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2836..2855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3069..3091
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3083..3100
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3084..3100
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3107..3126
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3107..3125
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3158..3174
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3158..3174
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 atatattaat atttattaaa atatataata tttagattat atataatata tataattttt
       61 atatatataa atatataaat tataaatata attattatta ttataatata ttagtgatta
      121 ccttatttat aaaggtatga agcaaggagt tattatttat tattattatt attattatta
      181 atataatata tatataatat attattaatt tttatttata ctccttcggg gtccccgccg
      241 ataagcttat cgataccgtc gacctcgagg gggggcccgg taccagcttt tgttcccttt
      301 agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      481 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      541 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      601 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      661 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1141 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1321 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1441 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1501 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1561 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1621 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1681 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1741 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1801 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1861 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1921 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1981 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2041 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2101 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2161 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2221 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2281 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2341 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2401 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2461 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     2521 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2581 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2641 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2701 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2761 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2821 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2881 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2941 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3001 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     3061 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact
     3121 atagggcgaa ttggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg
     3181 gctgcaggaa ttaattctca tgtttgacag cttatcatcg gggcccggcc acgggagccg
     3241 gaaccccgaa aggagaaata ttataattag aataatatct ttttttatta atattattat
     3301 taattaattt taataaatta gaacctttaa gtgtattatg taaacctttt atatagattt
     3361 ctataagtgt ttttcataat tggaaatcaa tttctttacg actatatcat tttaatgaat
     3421 ataatttagg taaaataaca taatataaaa aatctattct attaaattct aatttaataa
     3481 ctccattttt atcaggttta gttaatttaa tagatgaatc tcaatctttt aataatttat
     3541 ttttaattaa taatggtgta ttatcaattg gttttcaatt atttaataca taagatttaa
     3601 tagcttttaa tgtattaata ctacgtttat gttgtgaaac attaaaacca caaatattac
     3661 gttttggaga tagatcaaat gaaccttcag cttcaataaa acctaaaatt caataatcat
     3721 ttattttatc atatggaatt tttaaattag gtataacttc ataattatta attgtatttt
     3781 taatatttaa taaatcttta ttattataag atataccatt taaattatta attatagcta
     3841 tttttcattt ataataacta taatatttaa ttgttaaaca aggatactta tcaaaaatag
     3901 ttacaatatt agaatataac catttctttt gtgaatttac taatataatt gttttaagta
     3961 gttcttcaaa attagaaggt atatttaata tatttctaat cttttctaaa catgctttat
     4021 catcaatatg tattctaaaa ccataaaaaa atcttaaata ttttttatca tttaatttaa
     4081 tataaaaact accatctcca tctgtaaatc ctactaatca tttataaaaa ttatctattt
     4141 ctaaattatt cttattatta tttttattat ctttttgaac aataattaat ttattattta
     4201 aagaagtatg ataacctctt ttaataatat ttatattttt aatataaggt gattgaatag
     4261 aatataaatc tatatcttta ttatatttaa gaatattatt ataattatta ttattattat
     4321 tatttttaat aattaaaaat attaataata agtaaatatt aattattgtt catttaatca
     4381 ttccaaaaat ttaggtaatg atactgcttc gatcttaatt ggcatatttg catgacctgt
     4441 cccacacaac tcagaacatg ccccatagaa gacaccttct ctttgaatta aagcagaaac
     4501 ttgatttaat ctaccaggag tagcatcaac tttaatacct aaacttggga tagcaaaatc
     4561 atgaataaca tcagcagctg ttacaacgaa tctaatatgt gtatctacag gtacaactat
     4621 agaagtatca gtatctaata atcttaattg tccttcttct aataattcat caggaataac
     4681 atatgattca aattcaacag tttcaccact atcattaata aaatctgaat attcatattt
     4741 tcaatatcat tgatatccaa tagctttaat agttatagct ggtgaaataa cttcatcaca
     4801 taaatataat aaaataaatg aagggaaagc aataattaat aaaattacag ctggaaaaat
     4861 tgttcaaata acttcaatag tttgtccatg tttaatatat ttatatgcaa taggattttt
     4921 tgaatatgtt ataacaattg tatataacat tcaagatact aaacctaaaa taactaataa
     4981 ataaaacata atattatcat gtaattctaa aataccttct tgatttggtg ttgctgaatc
     5041 ctgaaaataa catgcataag gtgttggtac atcattcata atgaatgttg ttaattgtaa
     5101 tcttaataaa tctaacattt taataaatct taacctttag actcttttgt ctatttataa
     5161 tatgttaata ctacttttaa ttaaaattta tttataattt attattatat attataaata
     5221 aaataaaaat attatattac caaaccccac ctttttttat taaaaataat taaaattata
     5281 tattattttt ataaataata tatataatat tattaatcat ataaatatat attttataaa
     5341 atactaaaaa taaaactcct tcggggttcg gtccccctcc cgttagtata gggaggggtc
     5401 cctcactcct tacggaattc ataaggtgat tgaatagaat ataaatctat atctttatta
     5461 tatttaagaa tattattata attattatta ttattattat ttttaataat taaaaatatt
     5521 aataataagt aaatattaac tcgagttaag ctaaaattct ttcacataat cttcaacctg
     5581 ttacaccatt aattgttact ctaaataata atgaaccatc aggattaatt aatctttcat
     5641 caataatttt attaccattt cataatgtac ctgttactgt aattttttta ccatcaaata
     5701 cagcaatacc ttcataaggt ctaccaaaat aatcaattat attaggtgtt acaccatcaa
     5761 ttactaatgt accataatgt aaaattactt taaaatgatg atcatctaca ggatatacta
     5821 ctttaaaaat tttttcaatt tgacctattt gatcacctga taaaccttca taaggaataa
     5881 ttacatgaat atcaattttt aaaccatttt cacctgataa tacaattctt tgaataggtg
     5941 ttactgatac acctaaattt tgaaataatg atgatacacc accttgttct aatacttgat
     6001 ctaaattata accagctgtt tgtcttcaat cacctacaaa atcttctaat gtaaatacca
     6061 tatgaataaa tcttaacctt tagactcttt tgtctattta taatatgtta atactacttt
     6121 taattaaaat ttatgaattc ccgcttcccg aaaactaata atattcatat cat
//
