LOCUS       sequence-344740-        7019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chloroplast-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 7019)
  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 7019)
  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: cp-DdCBE G1333-C
FEATURES             Location/Qualifiers
     source          1..7019
                     /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             1405..1470
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2231..2335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2403..2421
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2459..2481)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2576..2899)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2794..2816
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2808..2825
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2809..2825
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2826..2882
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2895..2911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2919..2935
                     /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(2943..2949,2950..2967,2968..2973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2988..3009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3715..3966
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4692..4813
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4729..4748)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4783..4802
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4862..4878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4886..4902
                     /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(4910..4916,4917..4934,4935..4940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4955..4976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5264..5852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5344..5363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6023..6814,6815..6883))
                     /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(6884..6988)
                     /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 cgcaatggct tcctctatgc tctcttccgc tactatggtt
     1201 gcctctccgg ctcaggccac tatggtcgct cctttcaacg gacttaagtc ctccgctgcc
     1261 ttcccagcca cccgcaaggc taacaacgac attacttcca tcacaagcaa cggcggaaga
     1321 gttaactgca tgcaggtgtg gcctccgatt ggaaagaaga agtttgagac tctctcttac
     1381 cttcctgacc ttaccgattc cgaagactac aaggaccatg atggtgatta taaggatcat
     1441 gacattgatt acaaagatga tgatgacaaa atggatattg ctgatcttcg aacattgggt
     1501 tattctcagc agcaacaaga gaaaatcaaa ccgaaagtcc gttcaaccgt agctcaacat
     1561 catgaagccc ttgttggtca tggatttact catgctcaca ttgttgctct aagtcagcac
     1621 cctgccgcgc taggcacagt tgccgtcaag tatcaagata tgatagctgc tttacctgaa
     1681 gcaacccacg aggcaatagt gggggttggg aagaggggtg caggcgccag ggcactggag
     1741 gcactcttaa cagtagctgg agaacttcgg ggaccaccat tgcagcttga tactggacaa
     1801 ctcttgaaga tcgcaaaaag agggggtgtg actgctgtgg aagctgttca tgcatggaga
     1861 aatgctttga cgggagcacc cctgaagaga cctccgtaag atgcttttct gtgactggtg
     1921 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1981 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2041 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2101 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2161 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2221 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2281 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2341 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     2401 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     2461 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2521 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     2581 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     2641 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     2701 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2761 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2821 ccagtgaatt cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc
     2881 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2941 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3001 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3061 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ggtctcacgg tctgactccg
     3241 gaacaggtgg tggcgattgc aagcaacggc ggcggcaaac aggctctaga gtcgatagta
     3301 gcacaattaa gcaggcctga ccctgcactt gctgctctta caaatgatca tcttgttgct
     3361 ctggcctgtt tgggtgggcg tcccgcgttg gatgctgtta agaaaggact tggtggttct
     3421 ccaactcctt atccaaatta tgcaaatgct ggtcatgtgg aaggccagtc cgccctattc
     3481 atgagagata atggtatctc tgaagggcta gtctttcata ataatccaga aggaacttgt
     3541 ggattttgcg ttaacatgac tgagacgctg ctaccggaaa atgccaaaat gacagttgtt
     3601 ccaccagagg gcgcaattcc tgtgaagaga ggagcaactg gtgaaacaaa agttttcacc
     3661 gggaactcta actctcctaa atcaccaacc aaaggtggct gcagtggagg atcaacaaac
     3721 ctctccgaca taattgagaa agaaactggg aaacagttgg tcattcaaga atctattttg
     3781 atgcttcctg aagaagttga agaggtgatt ggaaataagc ccgagtcaga tattcttgta
     3841 cacactgctt atgatgaaag cacagatgag aatgtgatgt tgctcaccag tgatgcacct
     3901 gagtacaagc catgggcttt agtaatccaa gactcaaatg gagagaacaa gataaagatg
     3961 ttataataac acccagcttt caggcctccc agctttcgtc cgtatcatcg gtttcgacaa
     4021 cgttcgtcaa gttcaatgca tcagtttcat tgcccacaca ccagaatcct actaagtttg
     4081 agtattatgg cattggaaaa gctgttttct tctatcattt gttctgcttg taatttactg
     4141 tgttctttca gtttttgttt tcggacatca aaatgcaaat ggatggataa gagttaataa
     4201 atgatatggt ccttttgttc attctcaaat tattattatc tgttgttttt actttaatgg
     4261 gttgaattta agtaagaaag gaactaacag tgtgatatta aggtgcaatg ttagacatat
     4321 aaaacagtct ttcacctctc tttggttatg tcttgaattg gtttgtttct tcacttatct
     4381 gtgtaatcaa gtttactatg agtctatgat caagtaatta tgcaatcaag ttaagtacag
     4441 tataggcttt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     4501 atccccacgc tgcagtgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4561 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4621 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4681 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4741 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4801 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4861 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4921 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4981 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5041 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5101 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5161 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5221 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5281 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5341 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5401 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5461 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5521 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5581 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5641 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5701 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5761 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5881 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5941 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6061 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6121 gggagggctt accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg
     6181 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6241 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6301 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6361 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6421 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6481 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6541 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6601 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6661 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6721 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6781 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6841 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6901 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6961 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
