LOCUS       sequence-248049-        5116 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 5116)
  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 5116)
  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-N. tabacum trnI/trnA
FEATURES             Location/Qualifiers
     source          1..5116
                     /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(1638..1640,1641..1643,1644..2357)
                     /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(1683..1704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1944..1963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2291..2312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3322..3339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3432..4020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3512..3531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4194..4988)
                     /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(4989..5093)
                     /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 ggccttgtac acaccgcccg tcacactatg ggagctggcc atgcccgaag
      421 tcgttacctt aaccgcaagg agggggatgc cgaaggcagg gctagtgact ggagtgaagt
      481 cgtaacaagg tagccgtact ggaaggtgcg gctggatcac ctccttttca gggagagcta
      541 atgcttgttg ggtattttgg tttgacactg cttcacaccc ccaaaaaaaa gaagggagct
      601 acgtctgagt taaacttgga gatggaagtc atctttcctt tctcgacggt gaagtaagac
      661 caagctcatg agcttattat cctaggtcgg aacaagttga taggaccccc ttttttacgt
      721 ccccatgttc cccccgtgtg gcgacatggg ggcgaaaaaa ggaaagagag ggatggggtt
      781 tctctcgctt ttggcatagc gggcccccag tgggaggctc gcacgacggg ctattagctc
      841 agtggtagag cgcgcccctg ataattgcgt cgttgtgcct gggctgtgag ggctctcagc
      901 cacatggata gttcaatgtg ctcatcggcg cctgaccctg agatgtggat catccaaggc
      961 acattagcat ggcgtactcc tcctgttcga accggggttt gaaaccaaac tcctcctcag
     1021 gaggatagat ggggcgattc gggtgagatc caatgtagat ccaactttcg attcactcgt
     1081 gggatccggg cggtccgggg gggaccacca cggctcctct cttctcgaga atccatacat
     1141 cccttatcag tgtatggaca gctatctctc gagcacaggt ttagcaatgg gaaaataaaa
     1201 tggagcacct aacaacgcat cttcacagac caagaactac gagatcgccc ctttcattct
     1261 ggggtgacgg agggatcgta ccattcgagc cgtttttttc ttgactcgaa atgggagcag
     1321 gtttgaaaaa ggatcttaga gtgtctaggg ttgggccagg aggctctctt aacgccttct
     1381 tttttcttct catcggagtt atttcacaaa gacttgccag ggtaaggaag aaggggggaa
     1441 caagcacact tggagagcgc agtacaacgg agagttgtat gctgcgttcg ggaaggatga
     1501 atcgctcccg aaaaggaatc tattgattct ctcccaattg gttggaccgt aggtgcgatg
     1561 atttacttca cgggcgagct ctctggttca agtccaggat ggcccagctg cgaattcgtt
     1621 taaactgcct tgtcttcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1681 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1741 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1801 ccgtgccctg gcccaccctc gtgaccacct tgacctacgg cgtgcagtgc ttcgcccgct
     1861 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1921 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1981 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2041 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaaggtc tatatcaccg
     2101 ccgacaagca gaagaacggc atcaaggtga acttcaagac ccgccacaac atcgaggacg
     2161 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2221 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     2281 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2341 acgagctgta caagtaagaa gacaagggag tttaaacaag cttctccact tggctcgggg
     2401 ggatatagct cagttggtag agctccgctc ttgcaattgg gtcgttgcga ttacgggttg
     2461 gatgtctaat tgtccaggcg gtaatgatag tatcttgtac ctgaaccggt ggctcacttt
     2521 ttctaagtaa tggggaagag gaccgaaacg tgccactgaa agactctact gagacaaaga
     2581 tgggctgtca agaacgtaga ggaggtagga tgggcagttg gtcagatcta gtatggatcg
     2641 tacatggacg gtagttggag tcggcggctc tcccagggtt ccctcatctg agatctctgg
     2701 ggaagaggat caagttggcc cttgcgaaca gcttgatgca ctatctccct tcaacccttt
     2761 gagcgaaatg cggcaaaaga aaaggaagga aaatccatgg accgacccca tcatctccac
     2821 cccgtaggaa ctacgagatc accccaagga cgccttcggc atccaggggt cacggaccga
     2881 ccatagaacc ctgttcaata agtggaacgc attagctgtc cgctctcagg ttgggcagtc
     2941 agggtcggag aagggcaatg actcattctt agttagaatg ggattccaac tcagcacctt
     3001 ttgagtgaga ttttgagaag agttgctctt tggagagcac agtacgatga aagttgtaag
     3061 ctgtgttcgg gggggagtta ttgtctatcg ttggcctcta tggtagaatc agtcggggga
     3121 cctgagaggc ggtggtttac cctgcggcgg atgtcagcgg ttcgagtccg cttatctcca
     3181 actcgtgaac ttagccgata caaagctctg ggcctcatgg gccttccttt cactgcccgc
     3241 tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc
     3301 gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa
     3361 gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3421 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3481 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3541 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3601 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3661 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3721 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3781 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3841 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3901 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3961 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4021 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4081 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4141 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttattagaaa
     4201 aattcatcca gcagacgata aaacgcaata cgctggctat ccggtgccgc aatgccatac
     4261 agcaccagaa aacgatccgc ccattcgccg cccagttctt ccgcaatatc acgggtggcc
     4321 agcgcaatat cctgataacg atccgccacg cccagacggc cgcaatcaat aaagccgcta
     4381 aaacggccat tttccaccat aatgttcggc aggcacgcat caccatgggt caccaccaga
     4441 tcttcgccat ccggcatgct cgctttcaga cgcgcaaaca gctctgccgg tgccaggccc
     4501 tgatgttctt catccagatc atcctgatcc accaggcccg cttccatacg ggtacgcgca
     4561 cgttcaatac gatgtttcgc ctgatgatca aacggacagg tcgccgggtc cagggtatgc
     4621 agacgacgca tggcatccgc cataatgctc actttttctg ccggcgccag atggctagac
     4681 agcagatcct gacccggcac ttcgcccagc agcagccaat cacggcccgc ttcggtcacc
     4741 acatccagca ccgccgcaca cggaacaccg gtggtggcca gccagctcag acgcgccgct
     4801 tcatcctgca gctcgttcag cgcaccgctc agatcggttt tcacaaacag caccggacga
     4861 ccctgcgcgc tcagacgaaa caccgccgca tcagagcagc caatggtctg ctgcgcccaa
     4921 tcatagccaa acagacgttc cacccacgct gccgggctac ccgcatgcag gccatcctgt
     4981 tcaatcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5041 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5101 ccccgaaaag tgccac
//
