LOCUS       sequence-344830-        4686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondrial-targeting DdCBE (split DddA, G1397-N) Golden Gate 
            destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4686)
  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 4686)
  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-N
FEATURES             Location/Qualifiers
     source          1..4686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(176..764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(256..275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(935..1726,1727..1795))
                     /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     1558..1577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1796..1900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1968..1986
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2024..2046)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2141..2464)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2146..2165
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2359..2381
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2373..2390
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2374..2390
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2391..2447
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2460..2476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2484..2500
                     /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(2508..2514,2515..2532,2533..2538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2553..2574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3322..3573
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4290..4411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4327..4346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4381..4400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4460..4476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4484..4500
                     /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(4508..4514,4515..4532,4533..4538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4553..4574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
       61 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      121 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      181 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      241 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      301 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      361 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      421 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      481 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      541 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      601 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      661 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      721 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      781 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      841 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      901 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1021 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc
     1081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1141 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1381 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1441 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1621 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1861 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1921 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1981 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2041 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2101 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     2161 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2221 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     2281 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     2341 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
     2401 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     2461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2761 atacggttat ccacagaatc aggggggtct cacggtctga ctccggaaca ggtggtggcg
     2821 attgcaagca acggcggcgg caaacaggct ctagagtcca tagtagcaca actatcaaga
     2881 ccagatcctg ctctggctgc attgaccaat gatcacttgg ttgctttagc ttgcctcggt
     2941 gggcgtccgg ccttggatgc cgttaagaaa ggtttgggag gatctggatc ctatgcactg
     3001 ggaccatatc aaattagtgc gccgcaatta cctgcctaca atggacaaac agtagggaca
     3061 ttctactacg tcaatgacgc aggtggcctt gaatctaaag ttttttcttc tggcggtcct
     3121 actccttatc caaactatgc taatgctggt catgtggagg gacagtcagc acttttcatg
     3181 agggacaatg gtattagcga aggtcttgta tttcataata accccgaggg cacatgtgga
     3241 ttttgtgtaa acatgactga aaccctatta ccagagaatg caaaaatgac tgtggtccct
     3301 ccagaaggct cgggaggttc aacaaatctg agcgacatta ttgagaagga aacggggaag
     3361 cagctggtga ttcaggaaag tatcttgatg ttgcccgagg aggttgaaga agtgatagga
     3421 aacaaacctg agtcggatat acttgttcat actgcttatg atgagtctac tgatgaaaat
     3481 gttatgcttc tcacaagtga tgcacctgaa tataaaccat gggctttagt tatccaagat
     3541 tcaaatgggg aaaacaagat caagatgcta tgataatcca ggcctcccag ctttcgtccg
     3601 tatcatcggt ttcgacaacg ttcgtcaagt tcaatgcatc agtttcattg cccacacacc
     3661 agaatcctac taagtttgag tattatggca ttggaaaagc tgttttcttc tatcatttgt
     3721 tctgcttgta atttactgtg ttctttcagt ttttgttttc ggacatcaaa atgcaaatgg
     3781 atggataaga gttaataaat gatatggtcc ttttgttcat tctcaaatta ttattatctg
     3841 ttgtttttac tttaatgggt tgaatttaag taagaaagga actaacagtg tgatattaag
     3901 gtgcaatgtt agacatataa aacagtcttt cacctctctt tggttatgtc ttgaattggt
     3961 ttgtttcttc acttatctgt gtaatcaagt ttactatgag tctatgatca agtaattatg
     4021 caatcaagtt aagtacagta taggctttgg gctctatggc ttctgaggcg gaaagaacca
     4081 gctggggctc tagggggtat ccccacgctg cagtgagcgg gactctgggg ttcgaaatga
     4141 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4201 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4261 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4321 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4381 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4441 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4501 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4561 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4621 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4681 cttccg
//
