LOCUS Exported 3419 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoChlo kit. ACCESSION . VERSION . KEYWORDS pMK-Lux-E codon opt. Nt chloroplasts SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3419) 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 3419) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3419 /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" RBS 989..997 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" primer_bind complement(1625..1642) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1735..2323) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1815..1834) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2497..3291) /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(3292..3396) /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 aggccgcatg gtctcaaatg tctactttat taaatattga tgctactgaa attaaagtat 421 ctactgaaat tgatgatatt atttttactt cttctccttt aactttatta tttgaagatc 481 aagaaaaaat tcaaaaagaa ttaattttag aatcttttca ttatcattat aatcataata 541 aagattataa atattattgt aatattcaag gagtagatga aaatattcaa tctattgatg 601 atattcctgt atttcctact tctatgttta aatattctag attacatact gctgatgaat 661 ctaatattga aaattggttt acttcttctg gaactaaagg agtaaaatct catattgcta 721 gagatagaca atctattgaa agattattag gatctgtaaa ttatggaatg aaatatttag 781 gagaatttca tgaacatcaa ttagaattag taaatatggg acctgataga ttttctgctt 841 ctaatgtatg gtttaaatat gtaatgtctt tagtacaatt attatatcct actactttta 901 ctgtagaaaa tgatgaaatt gattttgaac aaactatttt agctttaaaa gctattcaaa 961 gaaaaggaaa aggaatttgt ttaattggac ctccttattt tatttattta ttatgtcatt 1021 atatgaaaga acataatatt gaatttaatg ctggagctca tatgtttatt attactggag 1081 gaggatggaa aactaaacaa aaagaagctt taaatagaca agattttaat caattattaa 1141 tggaaacttt ttctttattt catgaatctc aaattagaga tatttttaat caagtagaat 1201 taaatacttg tttttttgaa gattctttac aaagaaaaca tgtacctcct tgggtatatg 1261 ctagagcttt agatcctgta actttaactc ctgtagaaga tggacaagaa ggattaatgt 1321 cttatatgga tgcttcttct acttcttatc ctacttttat tgtaactgat gatattggaa 1381 ttgtaagaca tttaaaagaa cctgatcctt ttcaaggaac tactgtagaa attgtaagaa 1441 gattaaatac tagagaacaa aaaggatgtt ctttatctat ggctacttct ttaaaataag 1501 ctttgagacc ctgggcctca tgggccttcc tttcactgcc cgctttccag tcgggaaacc 1561 tgtcgtgcca gctgcattaa catggtcata gctgtttcct tgcgtattgg gcgctctccg 1621 cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg 1681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 1741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 1801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 1861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 1921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 1981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 2041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 2101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 2161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 2221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 2281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 2341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 2401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 2461 ctaaagtata tatgagtaaa cttggtctga cagttattag aaaaattcat ccagcagacg 2521 ataaaacgca atacgctggc tatccggtgc cgcaatgcca tacagcacca gaaaacgatc 2581 cgcccattcg ccgcccagtt cttccgcaat atcacgggtg gccagcgcaa tatcctgata 2641 acgatccgcc acgcccagac ggccgcaatc aataaagccg ctaaaacggc cattttccac 2701 cataatgttc ggcaggcacg catcaccatg ggtcaccacc agatcttcgc catccggcat 2761 gctcgctttc agacgcgcaa acagctctgc cggtgccagg ccctgatgtt cttcatccag 2821 atcatcctga tccaccaggc ccgcttccat acgggtacgc gcacgttcaa tacgatgttt 2881 cgcctgatga tcaaacggac aggtcgccgg gtccagggta tgcagacgac gcatggcatc 2941 cgccataatg ctcacttttt ctgccggcgc cagatggcta gacagcagat cctgacccgg 3001 cacttcgccc agcagcagcc aatcacggcc cgcttcggtc accacatcca gcaccgccgc 3061 acacggaaca ccggtggtgg ccagccagct cagacgcgcc gcttcatcct gcagctcgtt 3121 cagcgcaccg ctcagatcgg ttttcacaaa cagcaccgga cgaccctgcg cgctcagacg 3181 aaacaccgcc gcatcagagc agccaatggt ctgctgcgcc caatcatagc caaacagacg 3241 ttccacccac gctgccgggc tacccgcatg caggccatcc tgttcaatca tactcttcct 3301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 3361 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac //