LOCUS       40924_28451        7077 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLM006-mCherry-fusion, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7077)
  AUTHORS   Mackinder LC, Meyer MT, Mettler-Altmann T, Chen VK, Mitchell MC, 
            Caspari O, Freeman Rosenzweig ES, Pallesen L, Reeves G, Itakura A, 
            Roth R, Sommer F, Geimer S, Muhlhaus T, Schroda M, Goodenough U, 
            Stitt M, Griffiths H, Jonikas MC.
  TITLE     A repeat protein links Rubisco to form the eukaryotic 
            carbon-concentrating organelle
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2016) In press
  PUBMED    27166422
REFERENCE   2  (bases 1 to 7077)
  AUTHORS   Mackinder LCM., Meyer MT, Mettler-Altmann T, Chen V, Mitchell M, 
            Caspari O, Freeman Rosenzweig E, Pallesen L, Reeves G, Itakura A, 
            Roth R, Sommer F, Geimer S, Muhlhaus T, Schroda M, Goodenough U, 
            Stitt M, Griffiths H, Jonikas MC.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-APR-2016) Department of Plant Biology, Carnegie 
            Institution of Washington, 260 Panama Street, Stanford, CA 94305, 
            USA
REFERENCE   3  (bases 1 to 7077)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7077)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-APR-2016) Department of Plant Biology, Carnegie Institution of 
            Washington, 260 Panama Street, Stanford, CA 94305, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(423..509,655..1566)
                     /codon_start=1
                     /product="APHVII"
                     /label=APHVII
                     /note="hygromycin resistance marker"
                     /protein_id="ANF29842.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPGPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     promoter        complement(1827..1845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2900..2916
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2917..2973
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2986..3002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3010..3026)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3034..3064)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3079..3100)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3160..3867
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYR"
     CDS             3880..3897
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     rep_origin      complement(5258..5846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6020..6877)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6878..6982)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaattcga tatcaagctt ctttcttgcg
      121 ctatgacact tccagcaaaa ggtagggcgg gctgcgagac ggcttcccgg cgctgcatgc
      181 aacaccgatg atgcttcgac cccccgaagc tccttcgggg ctgcatgggc gctccgatgc
      241 cgctccaggg cgagcgctgt ttaaatagcc aggcccccga ttgcaaagac attatagcga
      301 gctaccaaag ccatattcaa acacctagat cactaccact tctacacagg ccactcgagc
      361 ttgtgatcgc actccgctaa gggggcgcct cttcctcttc gtttcagtca caacccgcaa
      421 acatgacaca agaatccctg ttacttctcg accgtattga ttcggatgat tcctacgcga
      481 gcctgcggaa cgaccaggaa ttctgggagg tgagtcgacg agcaagcccg gcggatcagg
      541 cagcgtgctt gcagatttga cttgcaacgc ccgcattgtg tcgacgaagg cttttggctc
      601 ctctgtcgct gtctcaagca gcatctaacc ctgcgtcgcc gtttccattt gcagccgctg
      661 gcccgccgag ccctggagga gctcgggctg ccggtgccgc cggtgctgcg ggtgcccggc
      721 gagagcacca accccgtact ggtcggcgag cccggcccgg tgatcaagct gttcggcgag
      781 cactggtgcg gtccggagag cctcgcgtcg gagtcggagg cgtacgcggt cctggcggac
      841 gccccggtgc cggtgccccg cctcctcggc cgcggcgagc tgcggcccgg caccggagcc
      901 tggccgtggc cctacctggt gatgagccgg atgaccggca ccacctggcg gtccgcgatg
      961 gacggcacga ccgaccggaa cgcgctgctc gccctggccc gcgaactcgg ccgggtgctc
     1021 ggccggctgc acagggtgcc gctgaccggg aacaccgtgc tcacccccca ttccgaggtc
     1081 ttcccggaac tgctgcggga acgccgcgcg gcgaccgtcg aggaccaccg cgggtggggc
     1141 tacctctcgc cccggctgct ggaccgcctg gaggactggc tgccggacgt ggacacgctg
     1201 ctggccggcc gcgaaccccg gttcgtccac ggcgacctgc acgggaccaa catcttcgtg
     1261 gacctggccg cgaccgaggt caccgggatc gtcgacttca ccgacgtcta tgcgggagac
     1321 tcccgctaca gcctggtgca actgcatctc aacgccttcc ggggcgaccg cgagatcctg
     1381 gccgcgctgc tcgacggggc gcagtggaag cggaccgagg acttcgcccg cgaactgctc
     1441 gccttcacct tcctgcacga cttcgaggtg ttcgaggaga ccccgctgga tctctccggc
     1501 ttcaccgatc cggaggaact ggcgcagttc ctctgggggc cgccggacac cgcccccggc
     1561 gcctgataag gatccccgct ccgtgtaaat ggaggcgctc gttgatctga gccttgcccc
     1621 ctgacgaacg gcggtggatg gaagatactg ctctcaagtg ctgaagcggt agcttagctc
     1681 cccgtttcgt gctgatcagt ctttttcaac acgtaaaaag cggaggagtt ttgcaatttt
     1741 gttggttgta acgatcctcc gttgattttg gcctctttct ccatgggcgg gctgggcgta
     1801 tttgaagcgg gtaccagctt ttgttccctt tagtgagggt taattgcgac caatcgtcac
     1861 acgagccctc gtcagaaaca cgtctccgcc acgctctccc tctcacggcc gaccccgcag
     1921 cccttttgcc ctttcctagg ccaccgacag gacccaggcg ctctcagcat gcctcaacaa
     1981 cccgtactcg tgccagcggt gcccttgtgc tggtgatcgc ttggaagcgc atgcgaagac
     2041 gaaggggcgg agcaggcggc ctggctgttc gaagggctcg ccgccagttc gggtgccttt
     2101 ctccacgcgc gcctccacac ctaccgatgc gtgaaggcag gcaaatgctc atgtttgccc
     2161 gaactcggtg tccttaaaaa gccgcttctt gtcgtcgttc cgagacatgt tagcagatcg
     2221 cagtgccacc tttcctgacg cgctcggccc catattcgga cgcaattgtc atttgtagca
     2281 caattggagc aaatctggcg aggcagtagg cttttaagtt gcaaggcgag agagcaaagt
     2341 gggacgcggc gtgattattg gtatttacgc gacggcccgg cgcgttagcg gcccttcccc
     2401 caggccaggg acgattatgt atcaatattg ttgcgttcgg gcactcgtgc gagggctcct
     2461 gcgggctggg gagggggatc tgggaattgg aggtacgacc gagatggctt gctcgggggg
     2521 aggtttcctc gccgagcaag ccagggttag gtgttgcgct cttgacacgt tgtgcattct
     2581 aggaccccac tgctactcac aacaagccca gttaacgccc gtcagggcgc gtcagcgggt
     2641 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2701 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     2761 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     2821 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     2881 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacccgggga
     2941 tcctctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3001 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3061 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3121 cgctttgtta acggagatct gggtggctcc ggcggccgga tggtgagcaa gggcgaggag
     3181 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3241 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3301 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     3361 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     3421 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3481 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     3541 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     3601 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3661 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3721 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     3781 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     3841 accggcggca tggacgagct gtacagatct ggcggtggcc accaccatca ccaccacgga
     3901 gatctctaac tgcacgaatt ctcaggtcgg gacatccccg accaggggcg gcggggatgt
     3961 tgctgggccg atggaaagta gcaacccagc cagcggcttc cagcgcactc cagctgctca
     4021 cggttgcgac attgcgcgtg cacgcttgcg cgtccctcac tcggccagct tgtcgccgca
     4081 gacatcccta gcattgtgcg gactgcggtc gtcagttagc gtagtggcgg ggctcaaagc
     4141 gtgatgcagc tggtggctga ttgcatgtgc tacatatgct gttatgtttt gcatgaactt
     4201 cgatgcattg gatgctgggt gcacgcgttt gcatgtgttt gtgccggcat gctgccgtcg
     4261 tcggccgtac gtttacgttt ctgtgtgccg gggtctttat ttccgcctgc agaccgacgt
     4321 cggttccact agttctagat ggcagcagct ggaccgcctg taccatggag aagagcttta
     4381 cttgccggga tggccgattt cgctgattga tacgggatcg gagctcggag gctttcgcgc
     4441 taggggctag gcgaagggca gtggtgacca gggtcggtgt ggggtcggcc cacggtcaat
     4501 tagccacagg aggatcaggg ggaggtaggc acgtcgactt ggtttgcgac cccgcagttt
     4561 tggcggacgt gctgttgtag atgttagcgt gtgcgtgagc cagtggccaa cgtgccacac
     4621 ccattgagaa gaccaaccaa cttactggca atatctgcca atgccatact gcatgtaatg
     4681 gccaggccat gtgagagttt gccgtgcctg cgcgcgcccc gggggcgaag cttggtaccg
     4741 ctagcgtcag ctgaactcct gctgccacgc tgacagagac gcgcccatga cgtagcgccg
     4801 actgctgctc ctctgctcgt ggagctctga atctgctgcc ggctgccccc gcccccgctg
     4861 ctgaggctga atctactccg ctctccaccc cagtcccagg aagagaggcg catttacttc
     4921 gcacagacgt tacagcacac ccttgatcat catcagctgc tcttccctgc cgctgcaaca
     4981 cgccccgcgc gatatcccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5041 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5101 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5161 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5221 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5281 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5341 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5401 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5461 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5521 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5581 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5641 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5701 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5761 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6001 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6061 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6121 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6901 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6961 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7021 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
