LOCUS       sequence-344742-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chloroplast-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 6827)
  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 6827)
  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 G1397-C
FEATURES             Location/Qualifiers
     source          1..6827
                     /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             3523..3774
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4500..4621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4537..4556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4591..4610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4670..4686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4694..4710
                     /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(4718..4724,4725..4742,4743..4748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4763..4784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5072..5660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5152..5171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5831..6622,6623..6691))
                     /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(6692..6796)
                     /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 gcaattcctg tgaagagagg agcaactggt gaaacaaaag ttttcaccgg gaactctaac
     3481 tctcctaaat caccaaccaa aggtggctgc agtggaggat caacaaacct ctccgacata
     3541 attgagaaag aaactgggaa acagttggtc attcaagaat ctattttgat gcttcctgaa
     3601 gaagttgaag aggtgattgg aaataagccc gagtcagata ttcttgtaca cactgcttat
     3661 gatgaaagca cagatgagaa tgtgatgttg ctcaccagtg atgcacctga gtacaagcca
     3721 tgggctttag taatccaaga ctcaaatgga gagaacaaga taaagatgtt ataataacac
     3781 ccagctttca ggcctcccag ctttcgtccg tatcatcggt ttcgacaacg ttcgtcaagt
     3841 tcaatgcatc agtttcattg cccacacacc agaatcctac taagtttgag tattatggca
     3901 ttggaaaagc tgttttcttc tatcatttgt tctgcttgta atttactgtg ttctttcagt
     3961 ttttgttttc ggacatcaaa atgcaaatgg atggataaga gttaataaat gatatggtcc
     4021 ttttgttcat tctcaaatta ttattatctg ttgtttttac tttaatgggt tgaatttaag
     4081 taagaaagga actaacagtg tgatattaag gtgcaatgtt agacatataa aacagtcttt
     4141 cacctctctt tggttatgtc ttgaattggt ttgtttcttc acttatctgt gtaatcaagt
     4201 ttactatgag tctatgatca agtaattatg caatcaagtt aagtacagta taggctttgg
     4261 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgctg
     4321 cagtgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4381 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4441 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4621 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4681 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4741 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4801 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4861 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4921 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4981 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5041 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5101 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5161 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5221 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5281 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5341 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5401 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5461 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5521 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5581 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5641 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5701 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5761 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5821 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5881 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5941 catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat
     6001 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6061 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6121 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6181 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6241 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6301 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6361 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6421 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6481 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6541 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6601 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6661 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6721 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6781 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
