LOCUS       sequence-348298-        4653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondrial-targeting DdCBE (split DddA, G1333-C) Golden Gate 
            destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4653)
  AUTHORS   Kang BC, Bae SJ, Lee S, Lee JS, Kim A, Lee H, Baek G, Seo H, Kim J, 
            Kim JS
  TITLE     Chloroplast and mitochondrial DNA editing in plants.
  JOURNAL   Nat Plants. 2021 Jul;7(7):899-905. doi: 10.1038/s41477-021-00943-9. 
            Epub 2021 Jul 1.
  PUBMED    34211132
REFERENCE   2  (bases 1 to 4653)
  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"; doi: 
            "10.1038/s41477-021-00943-9"; journalName: "Nat Plants"; date: 
            "2021-07"; volume: "7"; issue: "7"; pages: "899-905"
COMMENT     Sequence Label: mt-DdCBE G1333-C
FEATURES             Location/Qualifiers
     source          1..4653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1992..2010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2048..2070)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2165..2488)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2170..2189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2383..2405
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2397..2414
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2398..2414
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2415..2471
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2484..2500)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2508..2524
                     /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(2532..2538,2539..2556,2557..2562))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2577..2598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3304..3555
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4281..4402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4318..4337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4372..4391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4451..4467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4475..4491
                     /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(4499..4505,4506..4523,4524..4529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4544..4565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1981 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     2041 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2101 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     2161 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     2221 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     2281 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     2341 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     2401 aaacgacggc cagtgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg
     2461 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2521 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2581 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2641 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2701 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2761 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcaggggg gtctcacggt
     2821 ctgactccgg aacaggtggt ggcgattgca agcaacggcg gcggcaaaca ggctctagag
     2881 tcgatagtag cacaattaag caggcctgac cctgcacttg ctgctcttac aaatgatcat
     2941 cttgttgctc tggcctgttt gggtgggcgt cccgcgttgg atgctgttaa gaaaggactt
     3001 ggtggttctc caactcctta tccaaattat gcaaatgctg gtcatgtgga aggccagtcc
     3061 gccctattca tgagagataa tggtatctct gaagggctag tctttcataa taatccagaa
     3121 ggaacttgtg gattttgcgt taacatgact gagacgctgc taccggaaaa tgccaaaatg
     3181 acagttgttc caccagaggg cgcaattcct gtgaagagag gagcaactgg tgaaacaaaa
     3241 gttttcaccg ggaactctaa ctctcctaaa tcaccaacca aaggtggctg cagtggagga
     3301 tcaacaaacc tctccgacat aattgagaaa gaaactggga aacagttggt cattcaagaa
     3361 tctattttga tgcttcctga agaagttgaa gaggtgattg gaaataagcc cgagtcagat
     3421 attcttgtac acactgctta tgatgaaagc acagatgaga atgtgatgtt gctcaccagt
     3481 gatgcacctg agtacaagcc atgggcttta gtaatccaag actcaaatgg agagaacaag
     3541 ataaagatgt tataataaca cccagctttc aggcctccca gctttcgtcc gtatcatcgg
     3601 tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt gcccacacac cagaatccta
     3661 ctaagtttga gtattatggc attggaaaag ctgttttctt ctatcatttg ttctgcttgt
     3721 aatttactgt gttctttcag tttttgtttt cggacatcaa aatgcaaatg gatggataag
     3781 agttaataaa tgatatggtc cttttgttca ttctcaaatt attattatct gttgttttta
     3841 ctttaatggg ttgaatttaa gtaagaaagg aactaacagt gtgatattaa ggtgcaatgt
     3901 tagacatata aaacagtctt tcacctctct ttggttatgt cttgaattgg tttgtttctt
     3961 cacttatctg tgtaatcaag tttactatga gtctatgatc aagtaattat gcaatcaagt
     4021 taagtacagt ataggctttg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     4081 ctagggggta tccccacgct gcagtgagcg ggactctggg gttcgaaatg accgaccaag
     4141 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4201 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4261 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4321 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4381 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4441 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4501 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4561 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4621 attaatgaat cggccaacgc gcggggagag gcg
//
