LOCUS       sequence-248064-        5271 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 5271)
  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 5271)
  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-S. tuberosum trnI/trnA
FEATURES             Location/Qualifiers
     source          1..5271
                     /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             join(1645..1647,1648..1650,1651..2364)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1690..1711)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1951..1970)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2298..2319
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3477..3494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3587..4175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3667..3686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4349..5143)
                     /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(5144..5248)
                     /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 aggccgcatg tacacaccgc ccgtcacact atgggagctg gccatgcccg aagtcgttac
      421 cttaaccgca aggaggggga tgccgaaggc agggctagtg actggagtga agtcgtaaca
      481 aggtagccgt actggaaggt gcggctggat cacctccttt tcagggagag ctaatgcttg
      541 ttgggtattt tggtttgaca ctgcttcaca cccccaaaaa aaagaaggga gctacgtctg
      601 agttaaactt ggagatggaa gtcgtctttc ctttctcgac ggtgaagtaa gaccaagctc
      661 atgagcttat tatcctaggt cggaacaagt tgataggatc ccctttttta cgtccctatg
      721 ttcccccgtg tggcgacatg ggggcgaaaa aaggaaagag agggatgggg tttctctcgc
      781 ttttggcata gcgggccccc agtgggaggc tcgcacgacg ggctattagc tcagtggtag
      841 agcgcgcccc tgataattgc gtcgttgtgc ctgggctgtg agggctctca gccacatgga
      901 tagttcaatg tgctcatcgg cgcctgaccc tgagatgtgg atcatccaag gcacattagc
      961 atggcgtact cctcctgttc gaaccggggt ttgaaaccaa actcctcctc aggaggatag
     1021 atggggcgat tcgggtgaga tccaatgtag atccaacttt cgattcactc gtgggatccg
     1081 ggcggtccgg gggggaccac cacggctcct ctcttctcga gaatccatac atcccttatc
     1141 agtgtatgga cagctatctc tcgagcacag gtttaggttc ggcctcaatg ggaaaataaa
     1201 atggagcacc taacaacgca tcttcacaga ccaagaacta cgagatcacc cctttcattc
     1261 tggggtgacg gagggatcgt accattcgag ccgttttttt cttgactcga aatcgaaatg
     1321 ggagcaggtt tgaaaaagga tcttagagtg tctagggttg ggccaggagg atctcttaac
     1381 gccttctttt ttcttctcat cggagttatt tcacaaagac ttgccagggt aaggaagaag
     1441 gggggaacaa gcacacttgg agagcgcagt acaacggaga gttgtatgct gcgttcggga
     1501 aggatgaatc gctcccgaaa aggaatctat tgattctctc ccaattggtt ggaccgtagg
     1561 tgcgatgatt tacttcacgg gcgagatctc tggttcaagt ccaggatggc ccagctgcga
     1621 attcgtttaa actgccttgt cttcatggtg agcaagggcg aggagctgtt caccggggtg
     1681 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1741 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1801 aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc
     1861 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1921 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1981 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2041 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat
     2101 atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc
     2161 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2221 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     2281 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2341 ggcatggacg agctgtacaa gtaagaagac aagggagttt aaacaagctt ctccacttgg
     2401 ctcgggggga tatagctcag ttggtagagc tccgctcttg caattgggtc gttgcgatta
     2461 cgggttggat gtctaattgt ccaggcggta atgatagtat cttgtacctg aaccggtggc
     2521 tcactttttc taagtaatgg ggaagaggac cgaaacgtgc cactgaaaga ctctactgag
     2581 acaaagatgg gctgtcaaga acgtagagga ggtaggatgg gcagttggtc agatctagta
     2641 tggatcgtac atggacggta gttggagtcg gcggctctcc cagggttccc tcatctgaga
     2701 tctctgggga agaggatcaa gttggccctt gcgaacagct tgatgcacta tctcccttca
     2761 accctttgag cgaaatgcgg caaaagaaaa ggaaggaaaa tccatggacc gaccccatca
     2821 tctccacccc gtaggaacta cgagatcacc ccaaggacgc cttcggcatc caggggtcac
     2881 ggaccgacca tagaaccctg ttcaataagt ggaacgcatt agctgtccgc tctcaggttg
     2941 ggcagtcagg gtcggagaag ggcaatgact cattcttaaa accagcgttc ttaagaccaa
     3001 agagtcgggc ggaagggggg gaaagccctc cgttcctggt tctcctgtag ttggatcctc
     3061 cggaaccaca agaatcctta gttagaatgg gattccaact cagcaccttt tgagtgagat
     3121 tttgagaaga gttgctcttt ggagagcaca gtacgatgaa agttgtaagc tgtgttcggg
     3181 ggggagttat tgtctatcgt tggcctctat ggtagaatca gtcgggggac ctgagaggcg
     3241 gtggtttacc ctgcggcgga tgtcagcggt tcgagtccgc ttatctccaa ctcgtgaact
     3301 tagccgatac aaggctttat gatagcaccc aatttttccg attcggcggt tcgatctatg
     3361 atctgggcct catgggcctt cctttcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3421 cagctgcatt aacatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg
     3481 ctcactgact cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag
     3541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4021 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4141 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4201 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4261 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4321 tatatgagta aacttggtct gacagttatt agaaaaattc atccagcaga cgataaaacg
     4381 caatacgctg gctatccggt gccgcaatgc catacagcac cagaaaacga tccgcccatt
     4441 cgccgcccag ttcttccgca atatcacggg tggccagcgc aatatcctga taacgatccg
     4501 ccacgcccag acggccgcaa tcaataaagc cgctaaaacg gccattttcc accataatgt
     4561 tcggcaggca cgcatcacca tgggtcacca ccagatcttc gccatccggc atgctcgctt
     4621 tcagacgcgc aaacagctct gccggtgcca ggccctgatg ttcttcatcc agatcatcct
     4681 gatccaccag gcccgcttcc atacgggtac gcgcacgttc aatacgatgt ttcgcctgat
     4741 gatcaaacgg acaggtcgcc gggtccaggg tatgcagacg acgcatggca tccgccataa
     4801 tgctcacttt ttctgccggc gccagatggc tagacagcag atcctgaccc ggcacttcgc
     4861 ccagcagcag ccaatcacgg cccgcttcgg tcaccacatc cagcaccgcc gcacacggaa
     4921 caccggtggt ggccagccag ctcagacgcg ccgcttcatc ctgcagctcg ttcagcgcac
     4981 cgctcagatc ggttttcaca aacagcaccg gacgaccctg cgcgctcaga cgaaacaccg
     5041 ccgcatcaga gcagccaatg gtctgctgcg cccaatcata gccaaacaga cgttccaccc
     5101 acgctgccgg gctacccgca tgcaggccat cctgttcaat catactcttc ctttttcaat
     5161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
