LOCUS       sequence-344743-        6872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondrial-targeting DdCBE (split DddA, G1333-N) Golden Gate 
            destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6872)
  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 6872)
  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-N
FEATURES             Location/Qualifiers
     source          1..6872
                     /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             complement(1203..1214)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1396..1422
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1426..1452
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1456..1482
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2243..2347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2415..2433
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2471..2493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2588..2911)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2806..2828
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2820..2837
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2821..2837
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2838..2894
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2907..2923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2931..2947
                     /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(2955..2961,2962..2979,2980..2985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3000..3021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3577..3828
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4545..4666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4582..4601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4636..4655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4715..4731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4739..4755
                     /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(4763..4769,4770..4787,4788..4793))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4808..4829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5117..5705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5197..5216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5876..6667,6668..6736))
                     /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(6737..6841)
                     /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 cgcaatggca atggctgttt tccgtcgcga agggaggcgt
     1201 ctcctccctt caatcgccgc tcgcccaatc gctgctatcc gatctcctct ctcttctgac
     1261 caggaggaag gacttcttgg agttcgatct atctcaactc aagtggtgcg taaccgcatg
     1321 aagagtgtta agaacatcca aaagatcaca aaggcaatga agatggttgc tgcttccaag
     1381 cttagagcag ttcaatatcc ttacgatgtt ccagattacg ctggataccc ttatgacgtc
     1441 cctgattatg ctggttatcc atatgatgtg cccgactatg caatggatat agctgatttg
     1501 aggaccctcg ggtactcaca gcagcaacaa gaaaaaatca aaccaaaggt acgtagcacg
     1561 gttgcccaac atcatgaagc cctagttggt cacggattta ctcatgccca tattgttgca
     1621 ctgagtcaac accctgctgc attaggcaca gtggctgtga aatatcaaga tatgattgct
     1681 gctcttcctg aggccacaca cgaggcaatt gtgggtgttg gaaaacaatg gtctggtgcg
     1741 cgagcgcttg aagctctcct aaccgtagct ggagagttga gaggcccacc gcttcagctt
     1801 gacacagggc agctgttgaa gatagcaaag agggggggtg taactgctgt tgaagcagtc
     1861 catgcatgga gaaatgcatt aactggagct ccactgaaga gacctccgta agatgctttt
     1921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     2281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2341 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     2401 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     2461 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2521 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     2581 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     2641 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     2701 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2761 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2821 gtaaaacgac ggccagtgaa ttcgagctcg gtacccgggg atcctctaga gtcgacctgc
     2881 aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2941 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3001 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3061 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3121 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3181 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggggtctcac
     3241 ggtctgactc cggaacaggt ggtggcgatt gcaagcaacg gcggcggcaa acaggctcta
     3301 gagagcatag ttgctcagct aagtagacct gatccagcac ttgctgcatt gacgaacgat
     3361 catctcgtag cgctggcttg tttgggcggc aggccagcct tggacgccgt gaagaaagga
     3421 ttaggtggct cgggttcata cgctttaggg ccttatcaaa tttctgctcc tcagcttccc
     3481 gcatacaatg gtcaaactgt tggaacattt tactatgtca atgatgctgg tggtcttgag
     3541 tctaaagttt tctcctctgg tggaagtgga gggagcacca atctctccga catcattgag
     3601 aaagaaacag gaaagcagct ggtgatacaa gaatcaatat tgatgcttcc agaagaagtg
     3661 gaggaggtca ttggaaacaa gccggagtca gacattcttg tacacactgc ttatgatgaa
     3721 agtactgatg agaatgttat gttgttaaca tctgatgcac ctgaatataa accatgggca
     3781 ttagttattc aagattcaaa tggagaaaac aaaatcaaga tgctatgata atccaggcct
     3841 cccagctttc gtccgtatca tcggtttcga caacgttcgt caagttcaat gcatcagttt
     3901 cattgcccac acaccagaat cctactaagt ttgagtatta tggcattgga aaagctgttt
     3961 tcttctatca tttgttctgc ttgtaattta ctgtgttctt tcagtttttg ttttcggaca
     4021 tcaaaatgca aatggatgga taagagttaa taaatgatat ggtccttttg ttcattctca
     4081 aattattatt atctgttgtt tttactttaa tgggttgaat ttaagtaaga aaggaactaa
     4141 cagtgtgata ttaaggtgca atgttagaca tataaaacag tctttcacct ctctttggtt
     4201 atgtcttgaa ttggtttgtt tcttcactta tctgtgtaat caagtttact atgagtctat
     4261 gatcaagtaa ttatgcaatc aagttaagta cagtataggc tttgggctct atggcttctg
     4321 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgctgcagtg agcgggactc
     4381 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4441 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4501 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4561 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4621 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4681 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4741 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4801 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4861 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4921 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4981 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5041 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5101 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5161 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5221 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5281 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5341 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5401 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5461 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5521 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5581 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5641 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5701 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5761 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5821 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5881 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5941 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6001 ctgcaatgat accgcgagat ccacgctcac cggctccaga tttatcagca ataaaccagc
     6061 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6121 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6181 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6241 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6301 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6361 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6421 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6481 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6541 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6601 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6661 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6721 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6781 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6841 gcacatttcc ccgaaaagtg ccacctgacg tc
//
