LOCUS       sequence-344741-        7256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chloroplast-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 7256)
  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 7256)
  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-N
FEATURES             Location/Qualifiers
     source          1..7256
                     /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             1588..1614
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1618..1644
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1648..1674
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2435..2539)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2607..2625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2663..2685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2780..3103)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2998..3020
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3012..3029
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3013..3029
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3030..3086
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3099..3115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3123..3139
                     /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(3147..3153,3154..3171,3172..3177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3192..3213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3961..4212
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4929..5050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4966..4985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5020..5039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5099..5115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5123..5139
                     /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(5147..5153,5154..5171,5172..5177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5192..5213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5501..6089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5581..5600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6260..7051,7052..7120))
                     /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(7121..7225)
                     /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 cgcaatgctt caactctgct ccactttccg tcctcaactt
     1201 cttcttcctt gtcaattccg atttacaaat ggcgttttga ttccccaaat aaactatgtt
     1261 gcaagcaatt cagttgtgaa tatccggcca atgatacgat gccagagagc aagcggagga
     1321 agaggaggag ctaatagaag caaacctgct aagccgcaag ttaaagaagg aagcaacaaa
     1381 acagtaatcg aaggtttagt aaccgaatcg ctacctaatg gaatgtttcg agtcgatttg
     1441 gagaatggtg acaacattct tggttatatt tgtggtaaga ttaggaagaa tttcattcgg
     1501 atactccccg gtgataaagt gaaagtcgaa atgagtgttt atgattctac taaaggtcgt
     1561 atcattttca gaatgagtag tagagactat ccttacgatg ttccagatta cgctggatac
     1621 ccttatgacg tccctgatta tgctggttat ccatatgatg tgcccgacta tgcaatggat
     1681 atagctgatt tgaggaccct cgggtactca cagcagcaac aagaaaaaat caaaccaaag
     1741 gtacgtagca cggttgccca acatcatgaa gccctagttg gtcacggatt tactcatgcc
     1801 catattgttg cactgagtca acaccctgct gcattaggca cagtggctgt gaaatatcaa
     1861 gatatgattg ctgctcttcc tgaggccaca cacgaggcaa ttgtgggtgt tggaaaacaa
     1921 tggtctggtg cgcgagcgct tgaagctctc ctaaccgtag ctggagagtt gagaggccca
     1981 ccgcttcagc ttgacacagg gcagctgttg aagatagcaa agaggggggg tgtaactgct
     2041 gttgaagcag tccatgcatg gagaaatgca ttaactggag ctccactgaa gagacctccg
     2101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2581 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2641 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2701 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2761 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2821 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2881 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2941 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3001 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg ggatcctcta
     3061 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     3121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3241 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3301 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3361 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3421 ggggggtctc acggtctgac tccggaacag gtggtggcga ttgcaagcaa cggcggcggc
     3481 aaacaggctc tagagtccat agtagcacaa ctatcaagac cagatcctgc tctggctgca
     3541 ttgaccaatg atcacttggt tgctttagct tgcctcggtg ggcgtccggc cttggatgcc
     3601 gttaagaaag gtttgggagg atctggatcc tatgcactgg gaccatatca aattagtgcg
     3661 ccgcaattac ctgcctacaa tggacaaaca gtagggacat tctactacgt caatgacgca
     3721 ggtggccttg aatctaaagt tttttcttct ggcggtccta ctccttatcc aaactatgct
     3781 aatgctggtc atgtggaggg acagtcagca cttttcatga gggacaatgg tattagcgaa
     3841 ggtcttgtat ttcataataa ccccgagggc acatgtggat tttgtgtaaa catgactgaa
     3901 accctattac cagagaatgc aaaaatgact gtggtccctc cagaaggctc gggaggttca
     3961 acaaatctga gcgacattat tgagaaggaa acggggaagc agctggtgat tcaggaaagt
     4021 atcttgatgt tgcccgagga ggttgaagaa gtgataggaa acaaacctga gtcggatata
     4081 cttgttcata ctgcttatga tgagtctact gatgaaaatg ttatgcttct cacaagtgat
     4141 gcacctgaat ataaaccatg ggctttagtt atccaagatt caaatgggga aaacaagatc
     4201 aagatgctat gataatccag gcctcccagc tttcgtccgt atcatcggtt tcgacaacgt
     4261 tcgtcaagtt caatgcatca gtttcattgc ccacacacca gaatcctact aagtttgagt
     4321 attatggcat tggaaaagct gttttcttct atcatttgtt ctgcttgtaa tttactgtgt
     4381 tctttcagtt tttgttttcg gacatcaaaa tgcaaatgga tggataagag ttaataaatg
     4441 atatggtcct tttgttcatt ctcaaattat tattatctgt tgtttttact ttaatgggtt
     4501 gaatttaagt aagaaaggaa ctaacagtgt gatattaagg tgcaatgtta gacatataaa
     4561 acagtctttc acctctcttt ggttatgtct tgaattggtt tgtttcttca cttatctgtg
     4621 taatcaagtt tactatgagt ctatgatcaa gtaattatgc aatcaagtta agtacagtat
     4681 aggctttggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     4741 cccacgctgc agtgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4801 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4861 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4921 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4981 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5041 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5101 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5161 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5221 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5281 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5341 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5401 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5461 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5521 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5581 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5641 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5701 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5761 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5821 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5881 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5941 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6001 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6061 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6121 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6181 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6241 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6301 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6361 agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc
     6421 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6481 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6541 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6601 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6661 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6721 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6781 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6841 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6901 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6961 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7021 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7081 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7141 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7201 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
