LOCUS       sequence-338694-        6695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondrial-targeting DdCBE (split DddA, G1397-C) Golden Gate 
            destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6695)
  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 6695)
  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 G1397-C
FEATURES             Location/Qualifiers
     source          1..6695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(204..224)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             1273..1338
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2099..2203)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2271..2289
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2327..2349)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2444..2767)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2662..2684
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2676..2693
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2677..2693
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2694..2750
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2763..2779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2787..2803
                     /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(2811..2817,2818..2835,2836..2841))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2856..2877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3391..3642
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4368..4489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4405..4424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4459..4478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4538..4554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4562..4578
                     /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(4586..4592,4593..4610,4611..4616))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4631..4652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4940..5528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5020..5039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5699..6490,6491..6559))
                     /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"
     promoter        complement(6560..6664)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcc gttcaacttt tctatacaaa gttgaagatc tgactagcaa
      241 cgattgtaca attgcttctt taaaaaagga agaaagaaag aaagaaaaga atcaacatca
      301 gcgttaacaa acggccccgt tacggcccaa acggtcatat agagtaacgg cgttaagcgt
      361 tgaaagactc ctatcgaaat acgtaaccgc aaacgtgtca tagtcagatc ccctcttcct
      421 tcaccgcctc aaacacaaaa ataatcttct acagcctata tatacaaccc ccccttctat
      481 ctctcctttc tcacaattca tcatctttct ttctctaccc ccaattttaa gaaatcctct
      541 cttctcctct tcattttcaa ggtaaatctc tctctctctc tctctctctg ttattccttg
      601 ttttaattag gtatgtatta ttgctagttt gttaatctgc ttatcttatg tatgccttat
      661 gtgaatatct ttatcttgtt catctcatcc gtttagaagc tataaatttg ttgatttgac
      721 tgtgtatcta cacgtggtta tgtttatatc taatcagata tgaatttctt catattgttg
      781 cgtttgtgtg taccaatccg aaatcgttga tttttttcat ttaatcgtgt agctaattgt
      841 acgtatacat atggatctac gtatcaattg ttcatctgtt tgtgtttgta tgtatacaga
      901 tctgaaaaca tcacttctct catctgattg tgttgttaca tacatagata tagatctgtt
      961 atatcatttt ttttattaat tgtgtatata tatatgtgca tagatctgga ttacatgatt
     1021 gtgattattt acatgatttt gttatttacg tatgtatata tgtagatctg gactttttgg
     1081 agttgttgac ttgattgtat ttgtgtgtgt atatgtgtgt tctgatcttg atatgttatg
     1141 tatgtgcagc gaattcggcg cgcaatgttt aaacaagctt ctcgtctcct ctcccgatct
     1201 gtcgccgccg catcttccaa atcggtgacg actcgtgcct tttcaacgga acttccatcg
     1261 acgctcgatt ccgactacaa ggaccatgat ggtgattata aggatcatga cattgattac
     1321 aaagatgatg atgacaaaat ggatattgct gatcttcgaa cattgggtta ttctcagcag
     1381 caacaagaga aaatcaaacc gaaagtccgt tcaaccgtag ctcaacatca tgaagccctt
     1441 gttggtcatg gatttactca tgctcacatt gttgctctaa gtcagcaccc tgccgcgcta
     1501 ggcacagttg ccgtcaagta tcaagatatg atagctgctt tacctgaagc aacccacgag
     1561 gcaatagtgg gggttgggaa gaggggtgca ggcgccaggg cactggaggc actcttaaca
     1621 gtagctggag aacttcgggg accaccattg cagcttgata ctggacaact cttgaagatc
     1681 gcaaaaagag ggggtgtgac tgctgtggaa gctgttcatg catggagaaa tgctttgacg
     1741 ggagcacccc tgaagagacc tccgtaagat gcttttctgt gactggtgag tactcaacca
     1801 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1861 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1921 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1981 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2041 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2101 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2161 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2221 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     2281 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2341 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2401 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2461 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2521 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2581 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2641 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg
     2701 agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca
     2761 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2821 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2881 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2941 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3001 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3061 gtaatacggt tatccacaga atcagggggg tctcacggtc tgactccgga acaggtggtg
     3121 gcgattgcaa gcaacggcgg cggcaaacag gctctagagt cgatagtagc acaattaagc
     3181 aggcctgacc ctgcacttgc tgctcttaca aatgatcatc ttgttgctct ggcctgtttg
     3241 ggtgggcgtc ccgcgttgga tgctgttaag aaaggacttg gtggttctgc aattcctgtg
     3301 aagagaggag caactggtga aacaaaagtt ttcaccggga actctaactc tcctaaatca
     3361 ccaaccaaag gtggctgcag tggaggatca acaaacctct ccgacataat tgagaaagaa
     3421 actgggaaac agttggtcat tcaagaatct attttgatgc ttcctgaaga agttgaagag
     3481 gtgattggaa ataagcccga gtcagatatt cttgtacaca ctgcttatga tgaaagcaca
     3541 gatgagaatg tgatgttgct caccagtgat gcacctgagt acaagccatg ggctttagta
     3601 atccaagact caaatggaga gaacaagata aagatgttat aataacaccc agctttcagg
     3661 cctcccagct ttcgtccgta tcatcggttt cgacaacgtt cgtcaagttc aatgcatcag
     3721 tttcattgcc cacacaccag aatcctacta agtttgagta ttatggcatt ggaaaagctg
     3781 ttttcttcta tcatttgttc tgcttgtaat ttactgtgtt ctttcagttt ttgttttcgg
     3841 acatcaaaat gcaaatggat ggataagagt taataaatga tatggtcctt ttgttcattc
     3901 tcaaattatt attatctgtt gtttttactt taatgggttg aatttaagta agaaaggaac
     3961 taacagtgtg atattaaggt gcaatgttag acatataaaa cagtctttca cctctctttg
     4021 gttatgtctt gaattggttt gtttcttcac ttatctgtgt aatcaagttt actatgagtc
     4081 tatgatcaag taattatgca atcaagttaa gtacagtata ggctttgggc tctatggctt
     4141 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgctgca gtgagcggga
     4201 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4261 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4321 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4381 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4441 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4501 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4561 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4621 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4681 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4741 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4801 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4861 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4921 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4981 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5041 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5101 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5161 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5221 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5281 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5341 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5401 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5461 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5521 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5581 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5641 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5701 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5761 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5821 gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc
     5881 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5941 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6001 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6061 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6121 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6181 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6241 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6301 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6361 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6421 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6481 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6541 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6601 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6661 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
