LOCUS       sequence-248066-        5650 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 5650)
  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 5650)
  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 trnG/trnfM
FEATURES             Location/Qualifiers
     source          1..5650
                     /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(1558..1560,1561..1563,1564..2277)
                     /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(1603..1624)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1864..1883)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2211..2232
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3856..3873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3966..4554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4046..4065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4728..5522)
                     /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(5523..5627)
                     /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 cactaaagaa aaaaatctat tcaattagca aaaaaggaga gagagggatt
      421 cgaaccctcg atagttcttt gttcaaaact ataccggttt tcaagaccgg ggctatcaac
      481 cactcagcca tctctccgaa agactatttt tattttattc ctccgaatag aacatggcta
      541 taggggtgga tacccccact atctgtagaa agatctcagg tgcgaatcca ccggtcgatc
      601 tatctatccg tatatagata tatgatctag catgtccatt tgtgaaataa aaaagaaaat
      661 tccatttccc cccaccccac tccatgtacg aataaagtgc gaaaggggga gtagtaataa
      721 gtcatataga atcaatggat tcatgataaa gtaaaatccc tcgatgacat attttatcac
      781 aattaatatt ttttggctga tagagggatc aaatggtata tagttcattt gttggtagct
      841 tggaggatta aaagcatgac tcttgctttc caattggctg tttttgcatt aattgctact
      901 tcattaatct tattgattag cgtacccgtt gtatttgctt ctcctgatgg ctggtcaagt
      961 aacaaaaatg ttgtattttc tggtacatcc ttatggattg gattagtctt tctggtgggt
     1021 atccttaatt ctctcatctc ttgaacctat tcgtcgcaga cccaaaacca aaatgacccc
     1081 cctaattttt ctcggttgtg agacacatta aatttgaatc taagtcccca aagaaaatgc
     1141 aaatcaaata aagaaaacaa aaaaattaga ggggggtcaa acttcttgaa taaaaagaat
     1201 ataattaaca aaataattgg aatcgttccg aagagaatat gtgtcccggc actgcacaaa
     1261 taagatccgg ttatatatca tatatgtggg tacatattgt gtatcaagaa caaaaaaatg
     1321 cggatatggt cgaatggtaa aatttctctt tgccaaggag aagatgcggg ttcgattccc
     1381 gctatccgcc caagatccaa gataaagtaa tttttttact atttattttt attattgaat
     1441 ttcataaata gcattaaata tatccttaaa ttaaggagtt ggtatagttg gccgtgatag
     1501 tgtagtgatt ctatccctcc cctacgtttt cgaattcgtt taaactgcct tgtcttcatg
     1561 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1621 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1681 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1741 gtgaccacct tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     1801 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1861 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1921 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1981 ctggagtaca actacaacag ccacaaggtc tatatcaccg ccgacaagca gaagaacggc
     2041 atcaaggtga acttcaagac ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2101 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2161 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2221 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagaa
     2281 gacaagggag tttaaacaag cttcttacca gaataagatg ttgcggagac aggatttgaa
     2341 cccgtgacct caaggttatg agccttgcga gctaccaaac tgctctaccc cgcgccaaag
     2401 ataagaactg aaaactaata gataaacaag gattaaatgc gcccctccac cctatctgta
     2461 caaatagaat agcccattta tacagaatgg taaaggggct tctatgatca tcgaccatag
     2521 aaatagaaat gaagcgttaa tccttaccaa ctcgatcttg ttgctcctgg caacaaacat
     2581 gcatgaacca tttcacgaag tatgtgtccg gataggccaa agtctcgata gttagctctc
     2641 ggccttccgg tcaaaaaaca acgtcgatga aggcgtgtag gtgcactatt ccgtggtagg
     2701 gattgtaact ttccataaat ttcccatttg tcactcaacg acggaacctt gcttatttct
     2761 ttttttgagg atcgacgaat cgaatgatat ttctgttcca atttttgcct cttcttctcc
     2821 ctctgaatca aacttttcct tgccataatg gttgagttcc tattagtatc catgatacaa
     2881 gtcgaatcct agatgtagaa atagaagaag gtggaccccc ttctccgtcg aaagaaatga
     2941 gattatcgcg aatacacaca ttaattaacc aaatttgccc gacgtagagg caatcaagaa
     3001 agccgcataa gtgaatatat aacctacaga aaagtgagct aatccaacca atcttgcttg
     3061 tacaatagaa agggccactg gtttatctct ccatcgaatc aaattggcca aaggtgtgcg
     3121 ttcgtgagcc catgctaaag tttcaatcaa ttcctgccaa tatccacgcc aagaaattaa
     3181 gaacataaat ccagtagccc aaacaagatg tccaaataag aacatccatg cccaaaccga
     3241 taaactattc ataccaaaag ggttatatcc attgataagt tgtgaagagt ttaaccataa
     3301 ataatccctt aaccagccca tcaaataagt ggaagattca ttaaactgtg aaacgttacc
     3361 ttgccataat gtgatgtgct tccaatgcca ataaaaagta acccatccaa tagtatttaa
     3421 catccaaaaa actgccaaat aaaacgcgtc ccatgccgaa atatcacaag taccgcctcg
     3481 tcctgggcca tcgcacggaa aactataacc gaaatccttt ttatctggca ttaacttgga
     3541 accacgtgca tctaaagcac cttttactaa gatcaatgta gttgtatgta aaccaagggc
     3601 aatagcatga tgaaccaaaa agtctccagg acctattgtt aagaataatg aattactatt
     3661 ttcattaaca gcatttaacc aacccggcaa ccagatgctt cgacccgcat tgaatgctgg
     3721 gccagtcgtt gaagataaaa gctgggcctc atgggccttc ctttcactgc ccgctttcca
     3781 gtcgggaaac ctgtcgtgcc agctgcatta acatggtcat agctgtttcc ttgcgtattg
     3841 ggcgctctcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg taaagcctgg
     3901 ggtgcctaat gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3961 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4021 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4081 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4141 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4201 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4261 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4321 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4381 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4441 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4501 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4561 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4621 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4681 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttatta gaaaaattca
     4741 tccagcagac gataaaacgc aatacgctgg ctatccggtg ccgcaatgcc atacagcacc
     4801 agaaaacgat ccgcccattc gccgcccagt tcttccgcaa tatcacgggt ggccagcgca
     4861 atatcctgat aacgatccgc cacgcccaga cggccgcaat caataaagcc gctaaaacgg
     4921 ccattttcca ccataatgtt cggcaggcac gcatcaccat gggtcaccac cagatcttcg
     4981 ccatccggca tgctcgcttt cagacgcgca aacagctctg ccggtgccag gccctgatgt
     5041 tcttcatcca gatcatcctg atccaccagg cccgcttcca tacgggtacg cgcacgttca
     5101 atacgatgtt tcgcctgatg atcaaacgga caggtcgccg ggtccagggt atgcagacga
     5161 cgcatggcat ccgccataat gctcactttt tctgccggcg ccagatggct agacagcaga
     5221 tcctgacccg gcacttcgcc cagcagcagc caatcacggc ccgcttcggt caccacatcc
     5281 agcaccgccg cacacggaac accggtggtg gccagccagc tcagacgcgc cgcttcatcc
     5341 tgcagctcgt tcagcgcacc gctcagatcg gttttcacaa acagcaccgg acgaccctgc
     5401 gcgctcagac gaaacaccgc cgcatcagag cagccaatgg tctgctgcgc ccaatcatag
     5461 ccaaacagac gttccaccca cgctgccggg ctacccgcat gcaggccatc ctgttcaatc
     5521 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5581 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5641 aaagtgccac
//
