LOCUS       Exported                3017 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  MoChlo kit.
ACCESSION   .
VERSION     .
KEYWORDS    pMK-mEmerald
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3017)
  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 3017)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3017
                     /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             378..1097
                     /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(423..444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(684..703)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1031..1052
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1223..1240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1333..1921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1413..1432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2095..2889)
                     /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(2890..2994)
                     /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 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      421 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      481 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      541 ccgtgccctg gcccaccctc gtgaccacct tgacctacgg cgtgcagtgc ttcgcccgct
      601 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      661 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      721 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      781 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaaggtc tatatcaccg
      841 ccgacaagca gaagaacggc atcaaggtga acttcaagac ccgccacaac atcgaggacg
      901 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
      961 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     1021 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1081 acgagctgta caagtaagct ttgagaccct gggcctcatg ggccttcctt tcactgcccg
     1141 ctttccagtc gggaaacctg tcgtgccagc tgcattaaca tggtcatagc tgtttccttg
     1201 cgtattgggc gctctccgct tcctcgctca ctgactcgct gcgctcggtc gttcgggtaa
     1261 agcctggggt gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1741 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2041 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttattagaa
     2101 aaattcatcc agcagacgat aaaacgcaat acgctggcta tccggtgccg caatgccata
     2161 cagcaccaga aaacgatccg cccattcgcc gcccagttct tccgcaatat cacgggtggc
     2221 cagcgcaata tcctgataac gatccgccac gcccagacgg ccgcaatcaa taaagccgct
     2281 aaaacggcca ttttccacca taatgttcgg caggcacgca tcaccatggg tcaccaccag
     2341 atcttcgcca tccggcatgc tcgctttcag acgcgcaaac agctctgccg gtgccaggcc
     2401 ctgatgttct tcatccagat catcctgatc caccaggccc gcttccatac gggtacgcgc
     2461 acgttcaata cgatgtttcg cctgatgatc aaacggacag gtcgccgggt ccagggtatg
     2521 cagacgacgc atggcatccg ccataatgct cactttttct gccggcgcca gatggctaga
     2581 cagcagatcc tgacccggca cttcgcccag cagcagccaa tcacggcccg cttcggtcac
     2641 cacatccagc accgccgcac acggaacacc ggtggtggcc agccagctca gacgcgccgc
     2701 ttcatcctgc agctcgttca gcgcaccgct cagatcggtt ttcacaaaca gcaccggacg
     2761 accctgcgcg ctcagacgaa acaccgccgc atcagagcag ccaatggtct gctgcgccca
     2821 atcatagcca aacagacgtt ccacccacgc tgccgggcta cccgcatgca ggccatcctg
     2881 ttcaatcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2941 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3001 tccccgaaaa gtgccac
//
