LOCUS       sequence-365747-        5453 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 5453)
  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 5453)
  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
FEATURES             Location/Qualifiers
     source          1..5453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(171..190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      984..1572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1473..1492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1726..1743
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1860..1881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1896..1901,1902..1919,1920..1926)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1934..1950
                     /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     1939..1961
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1958..1974
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1958..1974
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1995..2013
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2041..2057
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2042..2058
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4588..4604)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4588..4604)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4636..4655)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4637..4655)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4662..4679)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4662..4678)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4671..4693)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4820..5275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4907..4926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5117..5138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5301..5405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5406..5453,1..21,22..813)
                     /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"
ORIGIN
        1 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
       61 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      121 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      181 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      241 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      301 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      361 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      421 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      481 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
      541 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
      601 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
      661 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
      721 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
      781 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
      841 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
      901 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
      961 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1021 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1081 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1141 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1201 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1261 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1321 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1381 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1441 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1501 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1561 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1621 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1681 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     1741 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     1801 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     1861 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     1921 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     1981 tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggtacc gggccccccc
     2041 tcgaggtcga cggtatcgat aagcttatcg gcggggaccc cgaaggagta taaataaaaa
     2101 ttaataatat attatatata tattatatta ataataataa taataataat aaataataac
     2161 tccttgcttc atacctttat aaataaggta atcactaata tattataata ataataatta
     2221 tatttataat ttatatattt atatatataa aaattatata tattatatat aatctaaata
     2281 ttatatattt taataaatat taatatatat gatatgaata ttattagttt tcgggaagcg
     2341 ggaattccgt aaggagtgag ggacccctcc ctatactaac gggaggggga ccgaaccccg
     2401 aaggagtttt atttttagta ttttataaaa tatatattta tatgattaat aatattatat
     2461 atattattta taaaaataat atataatttt aattattttt aataaaaaaa ggtggggttt
     2521 ggtaatataa tatttttatt ttatttataa tatataataa taaattataa ataaatttta
     2581 attaaaagta gtattaacat attataaata gacaaaagag tctaaaggtt aagatttatt
     2641 aaaatgttag atttattaag attacaatta acaacattca ttatgaatga tgtaccaaca
     2701 ccttatgcat gttattttca ggattcagca acaccaaatc aagaaggtat tttagaatta
     2761 catgataata ttatgtttta tttattagtt attttaggtt tagtatcttg aatgttatat
     2821 acaattgtta taacatattc aaaaaatcct attgcatata aatatattaa acatggacaa
     2881 actattgaag ttatttgaac aatttttcca gctgtaattt tattaattat tgctttccct
     2941 tcatttattt tattatattt atgtgatgaa gttatttcac cagctataac tattaaagct
     3001 attggatatc aatgatattg aaaatatgaa tattcagatt ttattaatga tagtggtaaa
     3061 ctgttgaatt ttgaatcata tgttattcct gatgaattat tagaagaagg acaattaaga
     3121 ttattagata ctgatacttc tatagttgta cctgtagata cacatattag attcgttgta
     3181 acagctgctg atgttattca tgattttgct atcccaagtt taggtattaa agttgatgct
     3241 actcctggta gattaaatca agtttctgct ttaattcaaa gagaaggtgt cttctatggg
     3301 gcatgttctg agttgtgtgg gacaggtcat gcaaatatgc caattaagat cgaagcagta
     3361 tcattaccta aatttttgga atgattaaat taacaataat taatatttac ttattattaa
     3421 tatttttaat tattaaaaat aataataata ataataatta taataatatt cttaaatata
     3481 ataaagatat agatttatat tctattcaat caccttatat taaaaatata aatattatta
     3541 aaagaggtta tcatacttct ttaaataata aattaattat tgttcaaaaa gataataaaa
     3601 ataataataa gaataattta gaaatagata atttttataa atgattagta ggatttacag
     3661 atggagatgg tagtttttat attaaattaa atgataaaaa atatttaaga tttttttatg
     3721 gttttagaat acatattgat gataaagcat gtttagaaaa gattagaaat atattaaata
     3781 taccttctaa ttttgaagaa ctacttaaaa caattatatt agtaaattca caaaagaaat
     3841 ggttatattc taatattgta actatttttg ataagtatcc ttgtttaaca attaaatatt
     3901 atagttatta taaatgaaaa atagctataa ttaataattt aaatggtata tcttataata
     3961 ataaagattt attaaatatt aaaaatacaa ttaataatta tgaagttata cctaatttaa
     4021 aaattccata tgataaaata aatgattatt gaattttagg ttttattgaa gctgaaggtt
     4081 catttgatct atctccaaaa cgtaatattt gtggttttaa tgtttcacaa cataaacgta
     4141 gtattaatac attaaaagct attaaatctt atgtattaaa taattgaaaa ccaattgata
     4201 atacaccatt attaattaaa aataaattat taaaagattg agattcatct attaaattaa
     4261 ctaaacctga taaaaatgga gttattaaat tagaatttaa tagaatagat tttttatatt
     4321 atgttatttt acctaaatta tattcattaa aatgatatag tcgtaaagaa attgatttcc
     4381 aattatgaaa aacacttata gaaatctata taaaaggttt acataataca cttaaaggtt
     4441 ctaatttatt aaaattaatt aataataata ttaataaaaa aagatattat tctaattata
     4501 atatttctcc tttcggggtt ccggctcccg tggccgggcc ccgatgataa gctgtcaaac
     4561 atgagaatta attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg
     4621 tggagctcca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     4681 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4741 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4801 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4861 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4921 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4981 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5041 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5101 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5161 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5221 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     5281 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5341 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5401 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gca
//
