LOCUS Exported 6004 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoChlo kit. ACCESSION . VERSION . KEYWORDS pMK-N. tabacum 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 attcaccgcc gtatggctga ccggcgatta ctagcgattc cggcttcatg 421 caggcgagtt gcagcctgca atccgaactg aggacgggtt tttggggtta gctcaccctc 481 gcgggatcgc gaccctttgt cccggccatt gtagcacgtg tgtcgcccag ggcataaggg 541 gcatgatgac ttgacgtcat cctcaccttc ctccggctta tcaccggcag tctgttcagg 601 gttccaaact caacgatggc aactaaacac gagggttgcg ctcgttgcgg gacttaaccc 661 aacaccttac ggcacgagct gacgacagcc atgcaccacc tgtgtccgcg ttcccgaagg 721 cacccctctc tttcaagagg attcgcggca tgtcaagccc tggtaaggtt cttcgctttg 781 catcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc ctttgagttt 841 cattcttgcg aacgtactcc ccaggcggga tacttaacgc gttagctaca gcactgcacg 901 ggtcgatacg cacagcgcct agtatccatc gtttacggct aggactactg gggtatctaa 961 tcccattcgc tcccctagct ttcctctctc agtgtcagtg tcggcccagc agagtgcttt 1021 cgccgttggt gttctttccg atctctacgc atttcaccgc tccaccggaa attccctctg 1081 cccctaccgt actccagctt ggtagtttcc accgcctgtc cagggttgag ccctgggatt 1141 tgacggcgga cttaaaaagc cacctacaga cgctttacgc ccaatcattc cggataacgc 1201 ttgcatcctc tgtattaccg cggctgctgg cacagagtta gccgatgctt attccccaga 1261 taccgtcatt gcttcttctc cgggaaaaga agttcacgac ccgtgggcct tctacctcca 1321 cgcggcattg ctccgtcagg ctttcgccca ttgcggaaaa ttccccactg ctgcctcccg 1381 taggagtctg ggccgtgtct cagtcccagt gtggctgatc atcctctcgg accagctact 1441 gatcatcgcc ttggtaagct attgcctcac caactagcta atcagacgcg agcccctcct 1501 cgggcggatt cctccttttg ctcctcagcc tacggggtat tagcagccgt ttccagctgt 1561 tgttcccctc ccaagggcag gttcttacgc gttactcacc cgtccgccac tggaaacacc 1621 acttcccgtc cgacttgcat gtgttaagca tgccgccagc gttcatcctg agccaggatc 1681 gaactctcca tgagattcat agttgcatta cttatagctt ccttgttcgt agacaaagcg 1741 gattcggaat tgtctttcat tccaaggcat aacttgtatc catgcgcttc atattcgccc 1801 ggagttcgct cccagaaata tagccatccc tgccccctca cgtcaatccc acgagcctct 1861 tatccattct cattgaacga cggcggggga gctttcgagg cctcgaaatc 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 ggtaacccta ccattaacta 3121 cttcatttat gaatttcata gtaatagaaa tacatgtcct accgagacag aatttggaac 3181 ttgctatcct cttgcctagc aggcaaagat ttacctccgt ggaaaggatg attcattcgg 3241 atcgacatga gagtccaact acattgccag aatccatgtt gtatatttga aagaggttga 3301 cctccttgct tctctcatgg tacactcctc ttcccgccga gccccttttc tcctcggtcc 3361 acagagacaa aatgtaggac tggtgccaac aattcatcag actcactaag tcgggatcac 3421 taactaatac taatctaata taatagtcta atatatctaa tataatagaa aatactaata 3481 taatagaaaa gaactgtctt ttctgtatac tttccccggt tccgttgcta ccgcgggctt 3541 tacgcaatcg atcggattag atagatatcc cttcaacata ggtcatcgaa aggatctcgg 3601 agagccacca aagtacgaaa gccaggatct ttcagaaaac ggattcctat tcaaagagtg 3661 cataaccgca tggataagct cacactaacc cgtcaatttg ggatccaaat tcgagatttt 3721 ccttgggagg tatcgggaag gatttggaat ggaataatat cgattcatac agaagaaaag 3781 gttctctatt gattcaaaca ctgtacctaa cctatgggat agggatcgag gaaggggaaa 3841 aaccgaagat ttcacatggt acttttatca atctgattta tttcgtacct ttcgttcaat 3901 gagaaaatgg gtcaaattct acaggatcaa acctatggga cttaaggaat gatataaaaa 3961 aaagagaggg aaaatattca tattaaataa atatgaagta gaagaaccca gattccaaat 4021 gaacaaattc aaacttgaaa aggatcttcc ttattcttga agaatgaggg gcaaagggat 4081 tgatcaagaa agatcctggg 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 //