LOCUS Exported 6004 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoChlo kit. ACCESSION . VERSION . KEYWORDS pMK-S. tuberosum trnV/3'rps12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6004) 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 6004) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6004 /organism="synthetic DNA construct" /mol_type="other DNA" 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 2185..2904 /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(2230..2251) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2491..2510) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2838..2859 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4210..4227) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4320..4908) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4400..4419) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5082..5876) /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(5877..5981) /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 acgagttcac cgccgtatgg ctgaccggcg attactagcg attccggctt 421 catgcaggcg agttgcagcc tgcaatccga actgaggacg ggtttttggg gttagctcac 481 cctcgcggga tcgcgaccct ttgtcccggc cattgtagca cgtgtgtcgc ccagggcata 541 aggggcatga tgacttgacg tcatcctcac cttcctccgg cttatcaccg gcagtctgtt 601 cagggttcca aactcaacga tggcaactaa acacgagggt tgcgctcgtt gcgggactta 661 acccaacacc ttacggcacg agctgacgac agccatgcac cacctgtgtc cgcgttcccg 721 aaggcacccc tctctttcaa gaggattcgc ggcatgtcaa gccctggtaa ggttcttcgc 781 tttgcatcga attaaaccac atgctccacc gcttgtgcgg gcccccgtca attcctttga 841 gtttcattct tgcgaacgta ctccccaggc gggatactta acgcgttagc tacagcactg 901 cacgggtcga tacgcacagc gcctagtatc catcgtttac ggctaggact actggggtat 961 ctaatcccat tcgctcccct agctttcggc tctcagtgtc agtgtcggcc cagcagagtg 1021 ctttcgccgt tggtgttctt tccgatctct acgcatttca ccgctccacc ggaaattccc 1081 tctgccccta ccgtactcca gcttggtagt ttccaccgcc tgtccagggt tgagccctgg 1141 gatttgacgg cggacttaaa aagccaccta cagacgcttt acgcccaatc attccggata 1201 acgcttgcat cctctgtatt accgcggctg ctggcacaga gttagccgat gcttattccc 1261 cagataccgt cattgcttct tctccgggaa aagaagttca cgacccgtgg gccttctacc 1321 tccacgcggc attgctccgt caggctttcg cccattgcgg aaaattcccc actgctgcct 1381 cccgtaggag tctgggccgt gtctcagtcc cagtgtggct gatcatcctc tcggaccagc 1441 tactgatcat cgccttggta agctattgcc tcaccaacta gctaatcaga cgcgagcccc 1501 tcctcgggcg gattcctcct tttgctcctc agcctacggg gtattagcag ccgtttccag 1561 ctgttgttcc cctcccaagg gcaggttctt acgcgttact cacccgtccg ccactggaaa 1621 caccgtttcc cgtgtttccc gtccgacttg catgtgttaa gcatgccgcc agcgttcatc 1681 ctgagccagg atcgaactct ccatgagatt catagttgca ttacttatag cttccttgtt 1741 cgtagacaaa gcggattcgg aattgtcttt cattccaagg cataacttgt atccatgcgc 1801 ttcatattcg cccggagttc gctcccagaa atatagtcat ccctgccccc tcacgtcaat 1861 cccacgagcc tcttatccat tctcattgaa cgacggcggg ggagcaaatc caactagaaa 1921 aactcacatt gggcttaggg ataatcaggc tcgaactgat gacttccacc acgtcaaggt 1981 gacactctac cgctgagtta tatcccttcc ccgccccatc gagaaataga actgactaat 2041 cctaagtcaa agggtcgaga aactcaacgc cactattctt gaacaacttg gagccgggcc 2101 ttcttttcgc actattacgg atatgaaaat aatggtcaaa atcggattca attgtcaaga 2161 attcgtttaa actgccttgt cttcatggtg agcaagggcg aggagctgtt caccggggtg 2221 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 2281 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 2341 aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc 2401 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 2461 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 2521 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 2581 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat 2641 atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc 2701 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 2761 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc 2821 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc 2881 ggcatggacg agctgtacaa gtaagaagac aagggagttt aaacaagctt ctgcccctat 2941 cggaaatagg attgactacc gattccgaag gaactggagt tacatctctt ttccattcaa 3001 gagttcttat gcgtttccac gcccctttaa gaccccgaaa aatggacaaa ttccttttct 3061 taggaacaca tacaagattc gtcactacaa aaaggataat gggaacccta ccattaacta 3121 cttcatttat gaatttcata gtaatagaaa tacatgtcct accgagacag aatttggaac 3181 ttgctaccct cttgcctagc aggcaaagat ttacctccgt ggaaaggatg attcattcgg 3241 atcgacatga gagtccaact acattgccag aatccatgtt gtatatttga aagaggttga 3301 cctccttgct tctctcatgg tacactcctc ttcccgccga gccccttttc tcctcggtcc 3361 acagagacaa aatgtaggac tggtgccaac agttcatcac ggaagaaagg actcactaag 3421 tcgggatcac taactaatac taatctaata taatagtcta atatatctaa tataatagaa 3481 aatactaata taatagaaaa gaactgtctt ttctgtatac tttccccggt tccgttgcta 3541 ccgcgggctt tacgcaatcg atcggattag atagatatcc cttcaacata ggtcatcgaa 3601 aggatctcgg atacccacca aagtacgaaa gccaggatct ttcagaaaac ggattcctat 3661 tcaaagagtg cataaccgca tggataagct cacactaacc cgtcaatttg ggatccaaat 3721 tagagatttt ccttgggagg tatcgggaag gatttggaat ggaataatat cgattcatac 3781 agaagaaaag gttctctatt gattcaaaca ctgtacctat gggataggga tcgaggaagg 3841 ggaaaaaccg aagatttcac atggtacttt tatcaatctg atttatttcg tacctttcgt 3901 tcaatgagaa aatgggtcaa attctacagg atcaaaccta tgggacttaa ggaatgatat 3961 aaaaaaaaga gagggaaaat attcatatta aataaatatg aagtagaaga acccagattc 4021 caaatgaaca aattcaaact tgaaaaggat cttccttatt cttgaagaat gaggggcaaa 4081 gggattgatc aagaactggg cctcatgggc cttcctttca ctgcccgctt tccagtcggg 4141 aaacctgtcg tgccagctgc attaacatgg tcatagctgt ttccttgcgt attgggcgct 4201 ctccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cgggtaaagc ctggggtgcc 4261 taatgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 4321 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 4381 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 4441 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 4501 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 4561 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 4621 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 4681 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 4741 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 4801 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 4861 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 4921 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 4981 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 5041 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt attagaaaaa ttcatccagc 5101 agacgataaa acgcaatacg ctggctatcc ggtgccgcaa tgccatacag caccagaaaa 5161 cgatccgccc attcgccgcc cagttcttcc gcaatatcac gggtggccag cgcaatatcc 5221 tgataacgat ccgccacgcc cagacggccg caatcaataa agccgctaaa acggccattt 5281 tccaccataa tgttcggcag gcacgcatca ccatgggtca ccaccagatc ttcgccatcc 5341 ggcatgctcg ctttcagacg cgcaaacagc tctgccggtg ccaggccctg atgttcttca 5401 tccagatcat cctgatccac caggcccgct tccatacggg tacgcgcacg ttcaatacga 5461 tgtttcgcct gatgatcaaa cggacaggtc gccgggtcca gggtatgcag acgacgcatg 5521 gcatccgcca taatgctcac tttttctgcc ggcgccagat ggctagacag cagatcctga 5581 cccggcactt cgcccagcag cagccaatca cggcccgctt cggtcaccac atccagcacc 5641 gccgcacacg gaacaccggt ggtggccagc cagctcagac gcgccgcttc atcctgcagc 5701 tcgttcagcg caccgctcag atcggttttc acaaacagca ccggacgacc ctgcgcgctc 5761 agacgaaaca ccgccgcatc agagcagcca atggtctgct gcgcccaatc atagccaaac 5821 agacgttcca cccacgctgc cgggctaccc gcatgcaggc catcctgttc aatcatactc 5881 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 5941 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 6001 ccac //