LOCUS       sequence-247992-        3932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoChlo kit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3932)
  AUTHORS   Occhialini A, Piatek A, Pfotenhauer AC, Frazier TP, Stewart CN, 
            Lenaghan S
  TITLE     MoChlo: A versatile modular cloning toolbox for chloroplast 
            biotechnology.
  JOURNAL   Plant Physiol. 2019 Jan 24. pii: pp.18.01220. doi: 
            10.1104/pp.18.01220.
  PUBMED    30679266
REFERENCE   2  (bases 1 to 3932)
  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"; journalName: "Plant 
            Physiol. 2019 Jan 24. pii: pp.18.01220. doi: 10.1104/pp.18.01220."
COMMENT     Sequence Label: pMK-Myc aadA-GFP
FEATURES             Location/Qualifiers
     source          1..3932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             343..354
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             425..454
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             461..1243
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="EAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSD
                     IDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRE
                     LQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEA
                     LNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVI
                     LEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(1310..1338)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1933..1953
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2138..2155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2248..2836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2328..2347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3010..3804)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(3805..3909)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca
      361 aggccacgtg tcttgtccgc ggtaccggtc tcaaatggca agcatgactg gtggacaggc
      421 tagcgaacaa aaactcattt ctgaagagga cttgcctagc gaagcggtga tcgccgaagt
      481 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
      541 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
      601 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
      661 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
      721 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
      781 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
      841 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
      901 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
      961 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     1021 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     1081 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     1141 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     1201 tgtccactac gtgaaaggcg agatcaccaa ggtagtgggc aaagaacttg cagttgaagg
     1261 aaaattggag gtcgccatgg ctagtaaagg agaagaactt ttcactggag ttgtcccaat
     1321 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     1381 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     1441 tgttccttgg ccaacacttg tcactacttt ctcttatggt gttcaatgct tttcaagata
     1501 cccagatcat atgaagcggc acgacttctt caagagcgcc atgcctgagg gatacgtgca
     1561 ggagaggacc atctctttca aggacgacgg gaactacaag acacgtgctg aagtcaagtt
     1621 tgagggagac accctcgtca acaggatcga gcttaaggga atcgatttca aggaggacgg
     1681 aaacatcctc ggccacaagt tggaatacaa ctacaactcc cacaacgtat acatcacggc
     1741 agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg
     1801 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     1861 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     1921 gagagatcac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     1981 tgaactatac aaataagctt tgagaccgag ctcggagcac aagactggcc tcatgggcct
     2041 tcctttcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taacatggtc
     2101 atagctgttt ccttgcgtat tgggcgctct ccgcttcctc gctcactgac tcgctgcgct
     2161 cggtcgttcg ggtaaagcct ggggtgccta atgagcaaaa ggccagcaaa aggccaggaa
     2221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3001 tgacagttat tagaaaaatt catccagcag acgataaaac gcaatacgct ggctatccgg
     3061 tgccgcaatg ccatacagca ccagaaaacg atccgcccat tcgccgccca gttcttccgc
     3121 aatatcacgg gtggccagcg caatatcctg ataacgatcc gccacgccca gacggccgca
     3181 atcaataaag ccgctaaaac ggccattttc caccataatg ttcggcaggc acgcatcacc
     3241 atgggtcacc accagatctt cgccatccgg catgctcgct ttcagacgcg caaacagctc
     3301 tgccggtgcc aggccctgat gttcttcatc cagatcatcc tgatccacca ggcccgcttc
     3361 catacgggta cgcgcacgtt caatacgatg tttcgcctga tgatcaaacg gacaggtcgc
     3421 cgggtccagg gtatgcagac gacgcatggc atccgccata atgctcactt tttctgccgg
     3481 cgccagatgg ctagacagca gatcctgacc cggcacttcg cccagcagca gccaatcacg
     3541 gcccgcttcg gtcaccacat ccagcaccgc cgcacacgga acaccggtgg tggccagcca
     3601 gctcagacgc gccgcttcat cctgcagctc gttcagcgca ccgctcagat cggttttcac
     3661 aaacagcacc ggacgaccct gcgcgctcag acgaaacacc gccgcatcag agcagccaat
     3721 ggtctgctgc gcccaatcat agccaaacag acgttccacc cacgctgccg ggctacccgc
     3781 atgcaggcca tcctgttcaa tcatactctt cctttttcaa tattattgaa gcatttatca
     3841 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3901 ggttccgcgc acatttcccc gaaaagtgcc ac
//
