LOCUS       sequence-248062-        5030 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 5030)
  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 5030)
  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 atpB/accD
FEATURES             Location/Qualifiers
     source          1..5030
                     /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(1376..1378,1379..1381,1382..2095)
                     /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(1421..1442)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1682..1701)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2029..2050
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3236..3253)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3346..3934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3426..3445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4108..4902)
                     /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(4903..5007)
                     /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 aggccgcatc actatctcga ccttgaacta ccagagcgtt ataaatattc ggcatcttgc
      421 ccgggggaaa ggctacatct agtaccggac cgatgatttg gacgacacgc cccgggtttt
      481 ttttttcaag cgtggaaacc ccagaaccag aagtagtagg attgattctc ataataataa
      541 aataaataaa tatgtcgaaa tgtttttgca aaaattatcg agttcaaaat aaatgtccgc
      601 tagcacgtcg atcggttaat tcaataaaat gggaattagc actcgatttc gttggcacca
      661 tgcaattgaa ccgattcaat tgtttactta ttcactgaga ctgagtgaat ttgcaagccc
      721 acccaaccta ttttaatttt aaaatctcaa gtggatgaat cagaatcttg agaaagtctt
      781 tcatttgtct atcattatag acaatcccat ccatattatc tattctatgg agttcgaacc
      841 tgaactttat tttctatttc tattacgatt cattatttgt atctaattgg ctcctcttct
      901 tatttatttt tgatttcaat ttcagcatat cgatttatgc ctagcctatt cttttctttg
      961 tgtttttctt tcttttttat acctttcata gattcataga ggagttccgt atattttcac
     1021 atctaggatt tacatataca acatatacca ctgtcaaggg ggaagttctt attatttagg
     1081 ttagtcaggt atttccattt caaaaaaaaa aaaagtaaaa aagaaaaatt gggttgcgct
     1141 atatatatga aagagtatac aataatgatg tatttggcaa atcaaatacc atggtctaat
     1201 aatcaaacat tctgattagt tgataatatt agtattagtt ggaaattttg tgaaagattc
     1261 ctgtgaaaag tttcattaac acggagttcg tgtcgagtag accttgttgt tgtgagagtt
     1321 cttaattcat gagttgtagg gagggatttg aattcgttta aactgccttg tcttcatggt
     1381 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1441 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1501 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1561 gaccaccttg acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1621 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1681 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1741 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1801 ggagtacaac tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat
     1861 caaggtgaac ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1921 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1981 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2041 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaagaaga
     2101 caagggagtt taaacaagct taaacagtag acattagcag ataaattagc aggaaataaa
     2161 gaaggataag gagaaagaac tcaagtaatt atccttcgtt ctcttaattg aattgcaatt
     2221 aaactcggcc caatctttta ctaaaaggat tgagccgaat acaacaaaga ttctattgca
     2281 tatattttga ctaagtatat acttacctag atatacaaga tttgaaatac aaaatctaga
     2341 aaactaaatc aaaatctaag actcaaatct ttctattgtt gtcttggatc cacaattaat
     2401 cctacggatc cttaggattg gtatattctt ttctatcctg tagtttgtag tttccctgaa
     2461 tcaagccaag tatcacacct ctttctaccc atcctgtata ttgtcccctt tgttccgtgt
     2521 tgaaatagaa ccttaattta ttacttattt ttttattaaa ttttagattt gttagtgatt
     2581 agatattagt attagacgag attttacgaa acaattattt ttttatttct ttataggaga
     2641 ggacaaatct cttttttcga tgcgaatttg acacgacata ggagaagccg ccctttatta
     2701 aaaattatat tattttaaat aatataaagg gggttccaac atattaatat atagtgaagt
     2761 gttcccccag attcagaact ttttttcaat actcacaatc cttattagtt aataatccta
     2821 gtgattggat ttctatgctt agtctgatag gaaataagat attcaaataa ataattttat
     2881 agcgaatgac tattcatcta ttgtattttc atgcaaatag ggggcaagaa aactctatgg
     2941 aaagatggtg gtttaattcg atgttgttta agaaggagtt cgaacgcagg tgtgggctaa
     3001 ataaatcaat gggcagtctt ggtcctattg aaaataccaa tgaagatcca aatcgaaaag
     3061 tgaaaaacat tcatagttgg aggaatcgtg acaattctag ttgcagtaat gttgattatt
     3121 tctgggcctc atgggccttc ctttcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3181 agctgcatta acatggtcat agctgtttcc ttgcgtattg ggcgctctcc gcttcctcgc
     3241 tcactgactc gctgcgctcg gtcgttcggg taaagcctgg ggtgcctaat gagcaaaagg
     3301 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3361 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3421 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3481 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3541 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3601 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3661 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3721 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3781 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3841 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3901 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3961 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4021 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4081 atatgagtaa acttggtctg acagttatta gaaaaattca tccagcagac gataaaacgc
     4141 aatacgctgg ctatccggtg ccgcaatgcc atacagcacc agaaaacgat ccgcccattc
     4201 gccgcccagt tcttccgcaa tatcacgggt ggccagcgca atatcctgat aacgatccgc
     4261 cacgcccaga cggccgcaat caataaagcc gctaaaacgg ccattttcca ccataatgtt
     4321 cggcaggcac gcatcaccat gggtcaccac cagatcttcg ccatccggca tgctcgcttt
     4381 cagacgcgca aacagctctg ccggtgccag gccctgatgt tcttcatcca gatcatcctg
     4441 atccaccagg cccgcttcca tacgggtacg cgcacgttca atacgatgtt tcgcctgatg
     4501 atcaaacgga caggtcgccg ggtccagggt atgcagacga cgcatggcat ccgccataat
     4561 gctcactttt tctgccggcg ccagatggct agacagcaga tcctgacccg gcacttcgcc
     4621 cagcagcagc caatcacggc ccgcttcggt caccacatcc agcaccgccg cacacggaac
     4681 accggtggtg gccagccagc tcagacgcgc cgcttcatcc tgcagctcgt tcagcgcacc
     4741 gctcagatcg gttttcacaa acagcaccgg acgaccctgc gcgctcagac gaaacaccgc
     4801 cgcatcagag cagccaatgg tctgctgcgc ccaatcatag ccaaacagac gttccaccca
     4861 cgctgccggg ctacccgcat gcaggccatc ctgttcaatc atactcttcc tttttcaata
     4921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
