LOCUS       40924_28426        7825 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLM005-RBCS1-Venus, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7825)
  AUTHORS   Freeman Rosenzweig ES, Xu B, Kuhn Cuellar L, Marinez-Sanchez A, 
            Schaffer M, Strauss M, Cartwright HN, Forster F, Engel BD, Wingreen 
            NS, Mackinder LCM., Jonikas MC.
  TITLE     The pyrenoid is liquid-like and exhibits dynamic reorganization
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7825)
  AUTHORS   Mackinder LCM., Jonikas MC.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JAN-2017) Department of Biology, University of York, 
            Wentworth Way, York, North Yorkshire YO10 5DD, United Kingdom
REFERENCE   3  (bases 1 to 7825)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7825)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JAN-2017) Department of Biology, University of York, Wentworth 
            Way, York, North Yorkshire YO10 5DD, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1256..1972
                     /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             2024..2047
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     rep_origin      complement(3408..3996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4170..5027)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5028..5132)
                     /label=AmpR promoter
     CDS             6011..6814
                     /codon_start=1
                     /product="APHVIII"
                     /label=APHVIII
                     /note="Paromomycin resistance marker"
                     /protein_id="AUR45269.1"
                     /translation="MDDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAA
                     LGAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRL
                     DVAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGER
                     LLAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLR
                     ELAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
     promoter        complement(7139..7157)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cggggggagg tttcctcgcc gagcaagcca gggttaggtg ttgcgctctt gacacgttgt
       61 gcattctagg accccactgc tactcacaac aagcccagtt atggccgccg tcattgccaa
      121 gtcctccgtc tccgcggccg tggctcgccc ggcccgctcc agcgtgcgcc ccatggccgc
      181 gctgaagccc gccgtcaagg ctgcccccgt ggctgccccg gctcaggcca accagatgat
      241 ggtctggacc ccggtcaaca acaagtgagt cgacgagcaa gcgcagcccg aaggataggg
      301 attctgcaag tcgcgacgca attgcttggg gcccagcctg ctgcctcaca tcgcacgtgc
      361 tctgccactt ctaacgagta ctggtcaatc gcgtgatcgc aggatgttcg agaccttctc
      421 ctacctgcct cctctgaccg acgagcagat cgccgcccag gtcgactaca tcgtcgccaa
      481 cggtgagcac ggcttggtgc cgtgggcata tgtagctggg ctgtgtttga gagctagtta
      541 atacgatcta gggcgtagct gcagactgcg acattacgat cgctcgaagc ttgagattag
      601 cttgttcgtt gtctgctgta tagtttgcct gtcatccgcc cttatctggg atagccctgg
      661 actgcttgcg tcggccactt atcttacccc cttcgtgctg tcgcttgcag gctggatccc
      721 ctgcctggag ttcgctgagg ccgacaaggc ctacgtgtcc aacgagtcgg ccatccgctt
      781 cggcagcgtg tcttgcgtaa gtcggagcga gagccagacg ggtccactgt ggcactgggt
      841 tagcttggca ccgggacagc gcctatctca ccgcggggaa ctgacgcata cccctgctcg
      901 tgcttcagca cggaaaagca aggggcccaa ttccatcttt ggtggttctg tgcgctggtg
      961 actgaacctc ttctccctcc catttcccgt gcgcccgcag ctgtactacg acaaccgcta
     1021 ctggaccatg tggaagctgc ccatgttcgg ctgccgcgac cccatgcagg tgctgcgcga
     1081 gatcgtcgcc tgcaccaagg ccttccccga tgcctacgtg cgcctggtgg ccttcgacaa
     1141 ccagaagcag gtgcagatca tgggcttcct ggtccagcgc cccaagactg cccgcgactt
     1201 ccagcccgcc aacaagcgct ccgtgaacgg agatctgggt ggctccggcg gccggatggt
     1261 gagcaagggc gaggagctgt tcaccggcgt ggtgcccatc ctggtggagc tggacggcga
     1321 cgtgaacggc cacaagttca gcgtgagcgg cgagggcgag ggcgacgcca cctacggcaa
     1381 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctggt
     1441 gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1501 cgacttcttc aagagcgcca tgcccgaggg ctacgtgcag gagcgcacca tcttcttcaa
     1561 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1621 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg gccacaagct
     1681 ggagtacaac tacaacagcc acaacgtgta catcaccgcc gacaagcaga agaacggcat
     1741 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tggccgacca
     1801 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1861 gagctaccag agcgccctga gcaaggaccc caacgagaag cgcgaccaca tggtgctgct
     1921 ggagttcgtg accgccgccg gcatcaccct gggcatggac gagctgatca agggcggtgg
     1981 cgactacaag gaccatgacg gtgactataa ggatcacgac atcgactaca aggacgatga
     2041 cgacaagtaa gatctctaac tgcacgaatt ctcaggtcgg gacatccccg accaggggcg
     2101 gcggggatgt tgctgggccg atggaaagta gcaacccagc cagcggcttc cagcgcactc
     2161 cagctgctca cggttgcgac attgcgcgtg cacgcttgcg cgtccctcac tcggccagct
     2221 tgtcgccgca gacatcccta gcattgtgcg gactgcggtc gtcagttagc gtagtggcgg
     2281 ggctcaaagc gtgatgcagc tggtggctga ttgcatgtgc tacatatgct gttatgtttt
     2341 gcatgaactt cgatgcattg gatgctgggt gcacgcgttt gcatgtgttt gtgccggcat
     2401 gctgccgtcg tcggccgtac gtttacgttt ctgtgtgccg gggtctttat ttccgcctgc
     2461 agaccgacgt cggttccact agttctagat ggcagcagct ggaccgcctg taccatggag
     2521 aagagcttta cttgccggga tggccgattt cgctgattga tacgggatcg gagctcggag
     2581 gctttcgcgc taggggctag gcgaagggca gtggtgacca gggtcggtgt ggggtcggcc
     2641 cacggtcaat tagccacagg aggatcaggg ggaggtaggc acgtcgactt ggtttgcgac
     2701 cccgcagttt tggcggacgt gctgttgtag atgttagcgt gtgcgtgagc cagtggccaa
     2761 cgtgccacac ccattgagaa gaccaaccaa cttactggca atatctgcca atgccatact
     2821 gcatgtaatg gccaggccat gtgagagttt gccgtgcctg cgcgcgcccc gggggcgaag
     2881 cttggtaccg ctagcgtcag ctgaactcct gctgccacgc tgacagagac gcgcccatga
     2941 cgtagcgccg actgctgctc ctctgctcgt ggagctctga atctgctgcc ggctgccccc
     3001 gcccccgctg ctgaggctga atctactccg ctctccaccc cagtcccagg aagagaggcg
     3061 catttacttc gcacagacgt tacagcacac ccttgatcat catcagctgc tcttccctgc
     3121 cgctgcaaca cgccccgcgc gatatcccag tcgggaaacc tgtcgtgcca gctgcattaa
     3181 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3241 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3301 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3361 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3421 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3841 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5161 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5221 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5281 acggtcacag cttgtctgta agcggatgcc gggagcagac aagatatcct tgacatgatt
     5341 ggtgcgtatg tttgtatgaa gctacaggac tgatttggcg ggctatgagg gcgggggaag
     5401 ctctggaagg gccgcgatgg ggcgcgcggc gtccagaagg cgccatacgg cccgctggcg
     5461 gcacccatcc ggtataaaag cccgcgaccc cgaacggtga cctccacttt cagcgacaaa
     5521 cgagcactta tacatacgcg actattctgc cgctatacat aaccactcag ctagcttaag
     5581 atcccatcaa gcttgcatgc cgggcgcgcc agaaggagcg cagccaaacc aggatgatgt
     5641 ttgatggggt atttgagcac ttgcaaccct tatccggaag ccccctggcc cacaaaggct
     5701 aggcgccaat gcaagcagtt cgcatgcagc ccctggagcg gtgccctcct gataaaccgg
     5761 ccagggggcc tatgttcttt acttttttac aagagaagtc actcaacatc ttaaaatggc
     5821 caggtgagtc gacgagcaag cccggcggat caggcagcgt gcttgcagat ttgacttgca
     5881 acgcccgcat tgtgtcgacg aaggcttttg gctcctctgt cgctgtctca agcagcatct
     5941 aaccctgcgt cgccgtttcc atttgcagga tggccactcc gccctccccg gtgctgaaga
     6001 atttcgaagc atggacgatg cgttgcgtgc actgcggggt cggtatcccg gttgtgagtg
     6061 ggttgttgtg gaggatgggg cctcgggggc tggtgtttat cggcttcggg gtggtgggcg
     6121 ggagttgttt gtcaaggtgg cagctctggg ggccggggtg ggcttgttgg gtgaggctga
     6181 gcggctggtg tggttggcgg aggtggggat tcccgtacct cgtgttgtgg agggtggtgg
     6241 ggacgagagg gtcgcctggt tggtcaccga agcggttccg gggcgtccgg ccagtgcgcg
     6301 gtggccgcgg gagcagcggc tggacgtggc ggtggcgctc gcggggctcg ctcgttcgct
     6361 gcacgcgctg gactgggagc ggtgtccgtt cgatcgcagt ctcgcggtga cggtgccgca
     6421 ggcggcccgt gctgtcgctg aagggagcgt cgacttggag gatctggacg aggagcggaa
     6481 ggggtggtcg ggggagcggc ttctcgccga gctggagcgg actcggcctg cggacgagga
     6541 tctggcggtt tgccacggtg acctgtgccc ggacaacgtg ctgctcgacc ctcgtacctg
     6601 cgaggtgacc gggctgatcg acgtggggcg ggtcggccgt gcggaccggc actccgatct
     6661 cgcgctggtg ctgcgcgagc tggcccacga ggaggacccg tggttcgggc cggagtgttc
     6721 cgcggcgttc ctgcgggagt acgggcgcgg gtgggatggg gcggtatcgg aggaaaagct
     6781 ggcgttttac cggctgttgg acgagttctt ctgagggacc tgatggtgtt ggtggctggg
     6841 tagggttgcg tcgcgtgggt gacagcacag tgtggacgtt gggatccccg ctccgtgtaa
     6901 atggaggcgc tcgttgatct gagccttgcc ccctgacgaa cggcggtgga tggaagatac
     6961 tgctctcaag tgctgaagcg gtagcttagc tccccgtttc gtgctgatca gtctttttca
     7021 acacgtaaaa agcggaggag ttttgcaatt ttgttggttg taacgatcct ccgttgattt
     7081 tggcctcttt ctccatgggc gggctgggcg tatttgaagc gggtaccagc ttttgttccc
     7141 tttagtgagg gttaattgcg accaatcgtc acacgagccc tcgtcagaaa cacgtctccg
     7201 ccacgctctc cctctcacgg ccgaccccgc agcccttttg ccctttccta ggccaccgac
     7261 aggacccagg cgctctcagc atgcctcaac aacccgtact cgtgccagcg gtgcccttgt
     7321 gctggtgatc gcttggaagc gcatgcgaag acgaaggggc ggagcaggcg gcctggctgt
     7381 tcgaagggct cgccgccagt tcgggtgcct ttctccacgc gcgcctccac acctaccgat
     7441 gcgtgaaggc aggcaaatgc tcatgtttgc ccgaactcgg tgtccttaaa aagccgcttc
     7501 ttgtcgtcgt tccgagacat gttagcagat cgcagtgcca cctttcctga cgcgctcggc
     7561 cccatattcg gacgcaattg tcatttgtag cacaattgga gcaaatctgg cgaggcagta
     7621 ggcttttaag ttgcaaggcg agagagcaaa gtgggacgcg gcgtgattat tggtatttac
     7681 gcgacggccc ggcgcgttag cggcccttcc cccaggccag ggacgattat gtatcaatat
     7741 tgttgcgttc gggcactcgt gcgagggctc ctgcgggctg gggaggggga tctgggaatt
     7801 ggaggtacga ccgagatggc ttgct
//
