LOCUS       Exported                3122 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  MoChlo kit.
ACCESSION   .
VERSION     .
KEYWORDS    pMK-aadA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3122)
  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 3122)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3122
                     /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             394..1185
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MAEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(1328..1345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1438..2026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1518..1537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2200..2994)
                     /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(2995..3099)
                     /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 aggccacgtg tcttgtccag agctcggtct caaatggcag aagcggtgat cgccgaagta
      421 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
      481 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
      541 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
      601 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
      661 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
      721 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
      781 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
      841 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
      901 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
      961 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     1021 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     1081 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     1141 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataagcttt gagaccggta
     1201 cctggagcac aagactggcc tcatgggcct tcctttcact gcccgctttc cagtcgggaa
     1261 acctgtcgtg ccagctgcat taacatggtc atagctgttt ccttgcgtat tgggcgctct
     1321 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg ggtaaagcct ggggtgccta
     1381 atgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1441 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1501 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1561 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1621 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1681 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1741 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1801 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1861 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1921 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1981 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2041 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2101 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2161 aatctaaagt atatatgagt aaacttggtc tgacagttat tagaaaaatt catccagcag
     2221 acgataaaac gcaatacgct ggctatccgg tgccgcaatg ccatacagca ccagaaaacg
     2281 atccgcccat tcgccgccca gttcttccgc aatatcacgg gtggccagcg caatatcctg
     2341 ataacgatcc gccacgccca gacggccgca atcaataaag ccgctaaaac ggccattttc
     2401 caccataatg ttcggcaggc acgcatcacc atgggtcacc accagatctt cgccatccgg
     2461 catgctcgct ttcagacgcg caaacagctc tgccggtgcc aggccctgat gttcttcatc
     2521 cagatcatcc tgatccacca ggcccgcttc catacgggta cgcgcacgtt caatacgatg
     2581 tttcgcctga tgatcaaacg gacaggtcgc cgggtccagg gtatgcagac gacgcatggc
     2641 atccgccata atgctcactt tttctgccgg cgccagatgg ctagacagca gatcctgacc
     2701 cggcacttcg cccagcagca gccaatcacg gcccgcttcg gtcaccacat ccagcaccgc
     2761 cgcacacgga acaccggtgg tggccagcca gctcagacgc gccgcttcat cctgcagctc
     2821 gttcagcgca ccgctcagat cggttttcac aaacagcacc ggacgaccct gcgcgctcag
     2881 acgaaacacc gccgcatcag agcagccaat ggtctgctgc gcccaatcat agccaaacag
     2941 acgttccacc cacgctgccg ggctacccgc atgcaggcca tcctgttcaa tcatactctt
     3001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3121 ac
//
