LOCUS       sequence-247995-        3893 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 3893)
  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 3893)
  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-FLARE16-S aadA-GFP
FEATURES             Location/Qualifiers
     source          1..3893
                     /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             407..1189
                     /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(1271..1299)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1894..1914
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2099..2116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2209..2797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2289..2308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2971..3765)
                     /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(3766..3870)
                     /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 tcaaatgggg gctagcgaag cggtgatcgc
      421 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
      481 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
      541 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
      601 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
      661 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
      721 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
      781 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
      841 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
      901 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
      961 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     1021 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     1081 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     1141 agaatttgtc cactacgtga aaggcgagat caccaaggta gtgggcaaag aacttgttga
     1201 aggaaaattg gagctagtag aaggtcttaa agtcgccatg gctagtaaag gagaagaact
     1261 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
     1321 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat
     1381 ttgcactact ggaaaactac ctgttccttg gccaacactt gtcactactt tctcttatgg
     1441 tgttcaatgc ttttcaagat acccagatca tatgaagcgg cacgacttct tcaagagcgc
     1501 catgcctgag ggatacgtgc aggagaggac catctctttc aaggacgacg ggaactacaa
     1561 gacacgtgct gaagtcaagt ttgagggaga caccctcgtc aacaggatcg agcttaaggg
     1621 aatcgatttc aaggaggacg gaaacatcct cggccacaag ttggaataca actacaactc
     1681 ccacaacgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat
     1741 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc
     1801 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     1861 ttcgaaagat cccaacgaaa agagagatca catggtcctt cttgagtttg taacagctgc
     1921 tgggattaca catggcatgg atgaactata caaataagct ttgagaccga gctcggagca
     1981 caagactggc ctcatgggcc ttcctttcac tgcccgcttt ccagtcggga aacctgtcgt
     2041 gccagctgca ttaacatggt catagctgtt tccttgcgta ttgggcgctc tccgcttcct
     2101 cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct aatgagcaaa
     2161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2881 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2941 tatatatgag taaacttggt ctgacagtta ttagaaaaat tcatccagca gacgataaaa
     3001 cgcaatacgc tggctatccg gtgccgcaat gccatacagc accagaaaac gatccgccca
     3061 ttcgccgccc agttcttccg caatatcacg ggtggccagc gcaatatcct gataacgatc
     3121 cgccacgccc agacggccgc aatcaataaa gccgctaaaa cggccatttt ccaccataat
     3181 gttcggcagg cacgcatcac catgggtcac caccagatct tcgccatccg gcatgctcgc
     3241 tttcagacgc gcaaacagct ctgccggtgc caggccctga tgttcttcat ccagatcatc
     3301 ctgatccacc aggcccgctt ccatacgggt acgcgcacgt tcaatacgat gtttcgcctg
     3361 atgatcaaac ggacaggtcg ccgggtccag ggtatgcaga cgacgcatgg catccgccat
     3421 aatgctcact ttttctgccg gcgccagatg gctagacagc agatcctgac ccggcacttc
     3481 gcccagcagc agccaatcac ggcccgcttc ggtcaccaca tccagcaccg ccgcacacgg
     3541 aacaccggtg gtggccagcc agctcagacg cgccgcttca tcctgcagct cgttcagcgc
     3601 accgctcaga tcggttttca caaacagcac cggacgaccc tgcgcgctca gacgaaacac
     3661 cgccgcatca gagcagccaa tggtctgctg cgcccaatca tagccaaaca gacgttccac
     3721 ccacgctgcc gggctacccg catgcaggcc atcctgttca atcatactct tcctttttca
     3781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
