LOCUS       sequence-248050-        6105 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 6105)
  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 6105)
  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 trnG/trnfM
FEATURES             Location/Qualifiers
     source          1..6105
                     /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(1571..1573,1574..1576,1577..2290)
                     /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(1616..1637)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1877..1896)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2224..2245
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4311..4328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4421..5009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4501..4520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5183..5977)
                     /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(5978..6082)
                     /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 aggccgcata ggagagagag ggattcgaac cctcgatagt tctttgttaa aactataccg
      421 gttttcaaga ccggggctat caaccgctca gccatctctc cgaaagacta tttttatttt
      481 attcctccga atagaacatg gccatagggg tggatacccc cactatctgt actatctgta
      541 aaaagatctc aggtgcgaat ccaccggtcg atctatctat ccgtatatag atatatgatc
      601 tagcatgccc atttgtgaaa taaaaaataa aattccattt ccccccactc catgtacgaa
      661 taaagtgcga aagggggagt agtaataagt catatagaat caatggattc atgataaagt
      721 aaaatccctc gatgacatat tttatcacaa ttaatatttt ttggctgata gagggatcaa
      781 atggtatata gttcatttgt tggtagcttg gaggattaaa agcatgactc ttgctttcca
      841 attggctgtt tttgcattaa ttgctacttc attaatctta ttgattagcg tacccgttgt
      901 atttgcttct cctgatggct ggtcaagtaa caaaaatgtt gtattttctg gtacatcctt
      961 atggattgga ttagtctttc tggtgggtat ccttaattct ctcatctctt gaacctattc
     1021 gtcgcagacc caaaaccaaa atgacccccc taatttttct cggttgtgag acacattaaa
     1081 ttggaatcta agtccccaaa gaaaacgcaa atcaaataaa gaaaacaaaa aaattagagg
     1141 ggggtcaaac ttcttgaata aaaagaatac aattaaaaaa ataattggaa tcgttccgaa
     1201 gagaatatgt gtcccggcac tgcacaaaaa agatccggtt atatatcata tatgtgggta
     1261 catattgtgt atcaagaaca aaaaaatgcg gatatggtcg aatggtaaaa tttctctttg
     1321 ccaaggagaa gatgcgggtt cgattcccgc tatccgccca agatccaaga taaagtaatt
     1381 ttattactat ttatttatta tttaatttca taaatagcat taaatatatc cttaaattaa
     1441 ggatttggta tagttggccg tgatagtgta gtgattctat ccctccccta cgttttcttt
     1501 ttccttccac ccccaaaaag cgaaaggcgg gaattaatta ctaggaattc gtttaaactg
     1561 ccttgtcttc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1621 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1681 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1741 ctggcccacc ctcgtgacca ccttgaccta cggcgtgcag tgcttcgccc gctaccccga
     1801 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1861 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1921 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1981 cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca ccgccgacaa
     2041 gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt
     2101 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2161 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     2221 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2281 gtacaagtaa gaagacaagg gagtttaaac aagcttaacc ctaaaatagt ttggcaaaac
     2341 aagatgttgc ggagacagga tttgaacccg tgacctcaag gttatgagcc ttgcgagcta
     2401 ccaaactgct ctaccccgcg ccgaagataa gaactgaaaa ctaatagata aacaaggatt
     2461 aaatgcgccc ctccacccta tctgtacaaa tagaatagcc catttataca gaatggtaaa
     2521 ggggcttcta tgatcatcga ccatagaaat agaaatgaag cgttaatcct taccaacttg
     2581 atcttgttgc tcctggcaac aaacatgcat gaaccatttc acgaagtatg tgtccggata
     2641 gtccaaagtc tcgatagtta gctctcggcc ttccggtcaa aaaacaacgt cgatgaaggc
     2701 gtgtaggtgc actattccgt ggtggggatt gtaactttcc ataaatttcc catttgtcac
     2761 tcaacgacgg aaccttgctt atttctttct ttgaggatcg acgaatcgaa tgatatttct
     2821 gttccaattt ttgcctcttc ttctccctct gaatcaaact tttccttgcc ataatggttg
     2881 agttcctatt agtatccatg atacaagtcg aatcctagat gtagaaatag aagaaggtgg
     2941 accccctctc cgtcgaaaga aatgagatta tcgcagatac acacattaaa aatattaacc
     3001 aaatttgccc gacgtagagg caatcaagaa agccgcataa gtgaatatat aacctacaga
     3061 aaagtgagct aatccaacca atcttgcttg tacaatggaa agggccactg gtttatctct
     3121 ccagcgaatc aaattggcca aaggtgtgcg ttcatgagcc catgctaaag tttcaatcaa
     3181 ttcctgccaa tatccacgcc aagaaattaa gaacataaat ccagtagccc aaacaagatg
     3241 tccaaataag aacatccatg cccaaaccga taaactattc ataccaaaag gattatatcc
     3301 gttgataagt tgtgaagagt ttaaccataa ataatccctt aaccagccca tcaaataagt
     3361 ggaagattca ttaaactgtg aaacgttacc ctgccataat gtgatgtgct tccaatgcca
     3421 ataaaaagta acccatccaa tagtatttaa catccaaaaa actgccaaat aaaacgcgtc
     3481 ccatgccgaa atatcacaag taccgcctcg tcctgggcca tcgcacggaa aactataacc
     3541 gaaatccttt ttatctggca ttaacttgga accacgtgca tctaaagcac cttttactaa
     3601 gatcaatgta gttgtatgta aaccaagagc aatagcatga tgaaccaaaa agtctccagg
     3661 acctattgtt aaaaataatg aattactatt ttcattaaca gcatttaacc aacccggcaa
     3721 ccagatgctt cgacccgcat tgaatgctgg accactcgtt gaagataaaa gtacatcgaa
     3781 cccatatgaa gttttaccat gagcggattg tatccattga gcaaatatag gttcaatcaa
     3841 gatttgcttc tccggagtgc caaaggcaag catgacatca ttatgaacat aaagtcccag
     3901 ggtatggaat cccagaaaga ggctggccca acttaaatga gatatgatag cttctttatg
     3961 ctctaacatt cttgccaata cattatcttc attttgctcc ggattgtaat ctctaatgaa
     4021 aaatatagct ccatgagcaa aagctcctgt catgatgaat cctgcgatat attggtggtg
     4081 ggtatataat gcagcttgag tagtaaagtc ttgtgctatg aatgcataag caggtaaaga
     4141 gtacatgtgt tgagctacca aagaagtaat aacccctaaa gaagctagag caaggcctgg
     4201 gcctcatggg ccttcctttc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4261 cattaacatg gtcatagctg tttccttgcg tattgggcgc tctccgcttc ctcgctcact
     4321 gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca aaaggccagc
     4381 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4921 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4981 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5041 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5101 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5161 agtaaacttg gtctgacagt tattagaaaa attcatccag cagacgataa aacgcaatac
     5221 gctggctatc cggtgccgca atgccataca gcaccagaaa acgatccgcc cattcgccgc
     5281 ccagttcttc cgcaatatca cgggtggcca gcgcaatatc ctgataacga tccgccacgc
     5341 ccagacggcc gcaatcaata aagccgctaa aacggccatt ttccaccata atgttcggca
     5401 ggcacgcatc accatgggtc accaccagat cttcgccatc cggcatgctc gctttcagac
     5461 gcgcaaacag ctctgccggt gccaggccct gatgttcttc atccagatca tcctgatcca
     5521 ccaggcccgc ttccatacgg gtacgcgcac gttcaatacg atgtttcgcc tgatgatcaa
     5581 acggacaggt cgccgggtcc agggtatgca gacgacgcat ggcatccgcc ataatgctca
     5641 ctttttctgc cggcgccaga tggctagaca gcagatcctg acccggcact tcgcccagca
     5701 gcagccaatc acggcccgct tcggtcacca catccagcac cgccgcacac ggaacaccgg
     5761 tggtggccag ccagctcaga cgcgccgctt catcctgcag ctcgttcagc gcaccgctca
     5821 gatcggtttt cacaaacagc accggacgac cctgcgcgct cagacgaaac accgccgcat
     5881 cagagcagcc aatggtctgc tgcgcccaat catagccaaa cagacgttcc acccacgctg
     5941 ccgggctacc cgcatgcagg ccatcctgtt caatcatact cttccttttt caatattatt
     6001 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6061 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
