LOCUS       40924_28436        7216 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLM005-Venus, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7216)
  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 7216)
  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 7216)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7216)
  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..7216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             784..1587
                     /codon_start=1
                     /product="APHVIII"
                     /label=APHVIII
                     /protein_id="ANF29834.1"
                     /translation="MDDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAA
                     LGAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRL
                     DVAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGER
                     LLAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLR
                     ELAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
     promoter        complement(1912..1930)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2985..3001
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3002..3058
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3071..3087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3095..3111)
                     /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(3119..3149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3164..3185)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3245..3961
                     /codon_start=1
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELIK"
     CDS             4013..4036
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     rep_origin      complement(5397..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6159..7016)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7017..7121)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagatat ccttgacatg attggtgcgt
      121 atgtttgtat gaagctacag gactgatttg gcgggctatg agggcggggg aagctctgga
      181 agggccgcga tggggcgcgc ggcgtccaga aggcgccata cggcccgctg gcggcaccca
      241 tccggtataa aagcccgcga ccccgaacgg tgacctccac tttcagcgac aaacgagcac
      301 ttatacatac gcgactattc tgccgctata cataaccact cagctagctt aagatcccat
      361 caagcttgca tgccgggcgc gccagaagga gcgcagccaa accaggatga tgtttgatgg
      421 ggtatttgag cacttgcaac ccttatccgg aagccccctg gcccacaaag gctaggcgcc
      481 aatgcaagca gttcgcatgc agcccctgga gcggtgccct cctgataaac cggccagggg
      541 gcctatgttc tttacttttt tacaagagaa gtcactcaac atcttaaaat ggccaggtga
      601 gtcgacgagc aagcccggcg gatcaggcag cgtgcttgca gatttgactt gcaacgcccg
      661 cattgtgtcg acgaaggctt ttggctcctc tgtcgctgtc tcaagcagca tctaaccctg
      721 cgtcgccgtt tccatttgca ggatggccac tccgccctcc ccggtgctga agaatttcga
      781 agcatggacg atgcgttgcg tgcactgcgg ggtcggtatc ccggttgtga gtgggttgtt
      841 gtggaggatg gggcctcggg ggctggtgtt tatcggcttc ggggtggtgg gcgggagttg
      901 tttgtcaagg tggcagctct gggggccggg gtgggcttgt tgggtgaggc tgagcggctg
      961 gtgtggttgg cggaggtggg gattcccgta cctcgtgttg tggagggtgg tggggacgag
     1021 agggtcgcct ggttggtcac cgaagcggtt ccggggcgtc cggccagtgc gcggtggccg
     1081 cgggagcagc ggctggacgt ggcggtggcg ctcgcggggc tcgctcgttc gctgcacgcg
     1141 ctggactggg agcggtgtcc gttcgatcgc agtctcgcgg tgacggtgcc gcaggcggcc
     1201 cgtgctgtcg ctgaagggag cgtcgacttg gaggatctgg acgaggagcg gaaggggtgg
     1261 tcgggggagc ggcttctcgc cgagctggag cggactcggc ctgcggacga ggatctggcg
     1321 gtttgccacg gtgacctgtg cccggacaac gtgctgctcg accctcgtac ctgcgaggtg
     1381 accgggctga tcgacgtggg gcgggtcggc cgtgcggacc ggcactccga tctcgcgctg
     1441 gtgctgcgcg agctggccca cgaggaggac ccgtggttcg ggccggagtg ttccgcggcg
     1501 ttcctgcggg agtacgggcg cgggtgggat ggggcggtat cggaggaaaa gctggcgttt
     1561 taccggctgt tggacgagtt cttctgaggg acctgatggt gttggtggct gggtagggtt
     1621 gcgtcgcgtg ggtgacagca cagtgtggac gttgggatcc ccgctccgtg taaatggagg
     1681 cgctcgttga tctgagcctt gccccctgac gaacggcggt ggatggaaga tactgctctc
     1741 aagtgctgaa gcggtagctt agctccccgt ttcgtgctga tcagtctttt tcaacacgta
     1801 aaaagcggag gagttttgca attttgttgg ttgtaacgat cctccgttga ttttggcctc
     1861 tttctccatg ggcgggctgg gcgtatttga agcgggtacc agcttttgtt ccctttagtg
     1921 agggttaatt gcgaccaatc gtcacacgag ccctcgtcag aaacacgtct ccgccacgct
     1981 ctccctctca cggccgaccc cgcagccctt ttgccctttc ctaggccacc gacaggaccc
     2041 aggcgctctc agcatgcctc aacaacccgt actcgtgcca gcggtgccct tgtgctggtg
     2101 atcgcttgga agcgcatgcg aagacgaagg ggcggagcag gcggcctggc tgttcgaagg
     2161 gctcgccgcc agttcgggtg cctttctcca cgcgcgcctc cacacctacc gatgcgtgaa
     2221 ggcaggcaaa tgctcatgtt tgcccgaact cggtgtcctt aaaaagccgc ttcttgtcgt
     2281 cgttccgaga catgttagca gatcgcagtg ccacctttcc tgacgcgctc ggccccatat
     2341 tcggacgcaa ttgtcatttg tagcacaatt ggagcaaatc tggcgaggca gtaggctttt
     2401 aagttgcaag gcgagagagc aaagtgggac gcggcgtgat tattggtatt tacgcgacgg
     2461 cccggcgcgt tagcggccct tcccccaggc cagggacgat tatgtatcaa tattgttgcg
     2521 ttcgggcact cgtgcgaggg ctcctgcggg ctggggaggg ggatctggga attggaggta
     2581 cgaccgagat ggcttgctcg gggggaggtt tcctcgccga gcaagccagg gttaggtgtt
     2641 gcgctcttga cacgttgtgc attctaggac cccactgcta ctcacaacaa gcccagttaa
     2701 cgcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     2761 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     2821 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     2881 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     2941 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     3001 tgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg
     3061 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3121 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3181 cattaattgc gttgcgctca ctgcccgctt tgttaacgga gatctgggtg gctccggcgg
     3241 ccggatggtg agcaagggcg aggagctgtt caccggcgtg gtgcccatcc tggtggagct
     3301 ggacggcgac gtgaacggcc acaagttcag cgtgagcggc gagggcgagg gcgacgccac
     3361 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     3421 caccctggtg accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat
     3481 gaagcagcac gacttcttca agagcgccat gcccgagggc tacgtgcagg agcgcaccat
     3541 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3601 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3661 ccacaagctg gagtacaact acaacagcca caacgtgtac atcaccgccg acaagcagaa
     3721 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     3781 ggccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3841 ccactacctg agctaccaga gcgccctgag caaggacccc aacgagaagc gcgaccacat
     3901 ggtgctgctg gagttcgtga ccgccgccgg catcaccctg ggcatggacg agctgatcaa
     3961 gggcggtggc gactacaagg accatgacgg tgactataag gatcacgaca tcgactacaa
     4021 ggacgatgac gacaagtaag atctctaact gcacgaattc tcaggtcggg acatccccga
     4081 ccaggggcgg cggggatgtt gctgggccga tggaaagtag caacccagcc agcggcttcc
     4141 agcgcactcc agctgctcac ggttgcgaca ttgcgcgtgc acgcttgcgc gtccctcact
     4201 cggccagctt gtcgccgcag acatccctag cattgtgcgg actgcggtcg tcagttagcg
     4261 tagtggcggg gctcaaagcg tgatgcagct ggtggctgat tgcatgtgct acatatgctg
     4321 ttatgttttg catgaacttc gatgcattgg atgctgggtg cacgcgtttg catgtgtttg
     4381 tgccggcatg ctgccgtcgt cggccgtacg tttacgtttc tgtgtgccgg ggtctttatt
     4441 tccgcctgca gaccgacgtc ggttccacta gttctagatg gcagcagctg gaccgcctgt
     4501 accatggaga agagctttac ttgccgggat ggccgatttc gctgattgat acgggatcgg
     4561 agctcggagg ctttcgcgct aggggctagg cgaagggcag tggtgaccag ggtcggtgtg
     4621 gggtcggccc acggtcaatt agccacagga ggatcagggg gaggtaggca cgtcgacttg
     4681 gtttgcgacc ccgcagtttt ggcggacgtg ctgttgtaga tgttagcgtg tgcgtgagcc
     4741 agtggccaac gtgccacacc cattgagaag accaaccaac ttactggcaa tatctgccaa
     4801 tgccatactg catgtaatgg ccaggccatg tgagagtttg ccgtgcctgc gcgcgccccg
     4861 ggggcgaagc ttggtaccgc tagcgtcagc tgaactcctg ctgccacgct gacagagacg
     4921 cgcccatgac gtagcgccga ctgctgctcc tctgctcgtg gagctctgaa tctgctgccg
     4981 gctgcccccg cccccgctgc tgaggctgaa tctactccgc tctccacccc agtcccagga
     5041 agagaggcgc atttacttcg cacagacgtt acagcacacc cttgatcatc atcagctgct
     5101 cttccctgcc gctgcaacac gccccgcgcg atatcccagt cgggaaacct gtcgtgccag
     5161 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5221 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5281 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5581 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5821 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6061 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6121 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6181 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6241 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6301 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6361 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6421 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6481 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6541 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6601 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6661 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6721 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6781 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6841 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6901 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6961 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7021 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7081 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7141 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7201 acgaggccct ttcgtc
//
