LOCUS       sequence-338692-        6887 bp    DNA     circular SYN 15-APR-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 6887)
  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 6887)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..6887
                     /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             3583..3834
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4560..4681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4597..4616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4651..4670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4730..4746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4754..4770
                     /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(4778..4784,4785..4802,4803..4808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4823..4844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5132..5720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5212..5231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5891..6682,6683..6751))
                     /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(6752..6856)
                     /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 gtggttctcc aactccttat
     3301 ccaaattatg caaatgctgg tcatgtggaa ggccagtccg ccctattcat gagagataat
     3361 ggtatctctg aagggctagt ctttcataat aatccagaag gaacttgtgg attttgcgtt
     3421 aacatgactg agacgctgct accggaaaat gccaaaatga cagttgttcc accagagggc
     3481 gcaattcctg tgaagagagg agcaactggt gaaacaaaag ttttcaccgg gaactctaac
     3541 tctcctaaat caccaaccaa aggtggctgc agtggaggat caacaaacct ctccgacata
     3601 attgagaaag aaactgggaa acagttggtc attcaagaat ctattttgat gcttcctgaa
     3661 gaagttgaag aggtgattgg aaataagccc gagtcagata ttcttgtaca cactgcttat
     3721 gatgaaagca cagatgagaa tgtgatgttg ctcaccagtg atgcacctga gtacaagcca
     3781 tgggctttag taatccaaga ctcaaatgga gagaacaaga taaagatgtt ataataacac
     3841 ccagctttca ggcctcccag ctttcgtccg tatcatcggt ttcgacaacg ttcgtcaagt
     3901 tcaatgcatc agtttcattg cccacacacc agaatcctac taagtttgag tattatggca
     3961 ttggaaaagc tgttttcttc tatcatttgt tctgcttgta atttactgtg ttctttcagt
     4021 ttttgttttc ggacatcaaa atgcaaatgg atggataaga gttaataaat gatatggtcc
     4081 ttttgttcat tctcaaatta ttattatctg ttgtttttac tttaatgggt tgaatttaag
     4141 taagaaagga actaacagtg tgatattaag gtgcaatgtt agacatataa aacagtcttt
     4201 cacctctctt tggttatgtc ttgaattggt ttgtttcttc acttatctgt gtaatcaagt
     4261 ttactatgag tctatgatca agtaattatg caatcaagtt aagtacagta taggctttgg
     4321 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgctg
     4381 cagtgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4441 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4501 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4561 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4621 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4681 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4741 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4801 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4861 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4921 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4981 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5041 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5101 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5161 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5221 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5281 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5341 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5401 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5461 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5521 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5581 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5641 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5701 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5761 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5821 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5881 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5941 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6001 catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat
     6061 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6121 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6181 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6241 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6301 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6361 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6421 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6481 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6541 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6601 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6661 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6721 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6781 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6841 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
