LOCUS       40924_33852        6698 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pOpt_mRuby2_Paro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6698)
  AUTHORS   Lauersen KJ, Mussgnug JH, Kruse O.
  TITLE     Efficient targeted expression of nuclear transgenes in Chlamydomonas
            reinhardtii with a novel, modular vector toolkit
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6698)
  AUTHORS   Lauersen KJ, Mussgnug JH, Kruse O.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2014) Algae Biotechnology and Bioenergy, 
            University of Bielefeld Center for Biotechnology, Universitaetsstr 
            27, Bielefeld, NRW 33615, Germany
REFERENCE   3  (bases 1 to 6698)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6698)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUN-2014) Algae Biotechnology and Bioenergy, University of 
            Bielefeld Center for Biotechnology, Universitaetsstr 27, Bielefeld, 
            NRW 33615, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     489..505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        515..533
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1247..1258
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             2296..2307
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             2320..2343
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     promoter        complement(4398..4416)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4437..4453)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4461..4477)
                     /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(4485..4515)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4530..4551)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4839..5427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5601..6458)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6459..6563)
                     /label=AmpR promoter
     rep_origin      complement(join(6590..6698,1..347))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
       61 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
      121 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
      181 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
      241 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
      301 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
      361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
      421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      481 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
      541 ggagctccac cgcggtggcg gccgctctag aactagtgct gaggcttgac atgattggtg
      601 cgtatgtttg tatgaagcta caggactgat ttggcgggct atgagggcgg gggaagctct
      661 ggaagggccg cgatggggcg cgcggcgtcc agaaggcgcc atacggcccg ctggcggcac
      721 ccatccggta taaaagcccg cgaccccgaa cggtgacctc cactttcagc gacaaacgag
      781 cacttataca tacgcgacta ttctgccgct atacataacc actcagctag cttaagatcc
      841 catcaccggt gcatgccggg cgcgccagaa ggagcgcagc caaaccagga tgatgtttga
      901 tggggtattt gagcacttgc aacccttatc cggaagcccc ctggcccaca aaggctaggc
      961 gccaatgcaa gcagttcgca tgcagcccct ggagcggtgc cctcctgata aaccggccag
     1021 ggggcctatg ttctttactt ttttacaaga gaagtcactc aacatcttaa aatggccagg
     1081 tgagtcgacg agcaagcccg gcggatcagg cagcgtgctt gcagatttga cttgcaacgc
     1141 ccgcattgtg tcgacgaagg cttttggctc ctctgtcgct gtctcaagca gcatctaacc
     1201 ctgcgtcgcc gtttccattt gcaggatgca tatgagatct gacgtcatcg agggcagggt
     1261 gagcaagggc gaggagctga tcaaggagaa catgcgcatg aaggtggtga tggagggcag
     1321 cgtgaacggc caccagttca agtgcaccgg cgagggcgag ggcaacccct acatgggcac
     1381 ccagaccatg cgcatcaagg tgatcgaggg cggccccctg cccttcgcct tcgacatcct
     1441 ggccaccagc ttcatgtacg gcagccgcac cttcatcaag taccccaagg gcatccccga
     1501 cttcttcaag cagagcttcc ccgagggctt cacctgggag cgcgtgaccc ggtacgagga
     1561 cggtgagctt gcggggttgc gagcaacact ccagcaacga acagtgccca agtcaggaat
     1621 ctgcagtcag cctgggcttt cggcggcttt ttcttgggca aacagcttgc actcatgcca
     1681 gcgcggcttg tccagcctca cttgagcttt ccagctgcta ccagccgggc tatacgacag
     1741 cgacagagcc atagcgtgga atcacttatt tgggttgccg aagtagcggt cggagcgtga
     1801 gttcttggtc aagccgcccc ttatccggtt cctgtccgtg tctttgtccc tcgttcaccc
     1861 ttcgcggcac ccttcatccc cttgcttgca ggtggcgtgg tgaccgtgat gcaggacacc
     1921 agcctggagg acggctgcct ggtgtaccac gtgcaggtgc gcggcgtgaa cttccccagc
     1981 aacggccccg tgatgcagaa gaagaccaag ggctgggagc ccaacaccga gatgatgtac
     2041 cccgccgacg gcggcctgcg cggctacacc cacatggccc tgaaggtgga cggcggcggc
     2101 cacctgagct gcagcttcgt gaccacctac cgcagcaaga agaccgtggg caacatcaag
     2161 atgcccggca tccacgccgt ggaccaccgc ctggagcgcc tggaggagag cgacaacgag
     2221 atgttcgtgg tgcagcgcga gcacgccgtg gccaagttcg ccggcctggg cggcggcatg
     2281 gacgagctgt acaagatcga gggcagggat atcgaattct ggagccaccc gcagttcgag
     2341 aagtaaccgc tccgtgtaaa tggaggcgct cgttgatctg agccttgccc cctgacgaac
     2401 ggcggtggat ggaagatact gctctcaagt gctgaagcgg tagcttagct ccccgtttcg
     2461 tgctgatcag tctttttcaa cacgtaaaaa gcggaggagt tttgcaattt tgttggttgt
     2521 aacgatcctc cgttgatttt ggcctctttc tccatgggcg ggctgggcgt atttgaagcg
     2581 ggatccttcg aacgtacggt taactcgcga cctgcaggta cgtaatttaa atacgcgtgc
     2641 tgaggcttga catgattggt gcgtatgttt gtatgaagct acaggactga tttggcgggc
     2701 tatgagggcg ggggaagctc tggaagggcc gcgatggggc gcgcggcgtc cagaaggcgc
     2761 catacggccc gctggcggca cccatccggt ataaaagccc gcgaccccga acggtgacct
     2821 ccactttcag cgacaaacga gcacttatac atacgcgact attctgccgc tatacataac
     2881 cactcagcta gcttaagatc ccatccctag ggcatgccgg gcgcgccaga aggagcgcag
     2941 ccaaaccagg atgatgtttg atggggtatt tgagcacttg caacccttat ccggaagccc
     3001 cctggcccac aaaggctagg cgccaatgca agcagttcgc atgcagcccc tggagcggtg
     3061 ccctcctgat aaaccggcca gggggcctat gttctttact tttttacaag agaagtcact
     3121 caacatctta aaatggccag gtgagtcgac gagcaagccc ggcggatcag gcagcgtgct
     3181 tgcagatttg acttgcaacg cccgcattgt gtcgacgaag gcttttggct cctctgtcgc
     3241 tgtctcaagc agcatctaac cctgcgtcgc cgtttccatt tgcaggaagc ttactccgcc
     3301 ctccccggtg ctgaagaatt tcgaagcatg gacgatgcgt tgcgtgcact gcggggtcgg
     3361 tatcccggtt gtgagtgggt tgttgtggag gatggggcct cgggggctgg tgtttatcgg
     3421 cttcggggtg gtgggcggga gttgtttgtc aaggtggcag ctctgggggc cggggtgggc
     3481 ttgttgggtg aggctgagcg gctggtgtgg ttggcggagg tggggattcc cgtacctcgt
     3541 gttgtggagg gtggtgggga cgagagggtc gcctggttgg tcaccgaagc ggttccgggg
     3601 cgtccggcca gtgcgcggtg gccgcgggag cagcggctgg acgtggcggt ggcgctcgcg
     3661 gggctcgctc gttcgctgca cgcgctggac tgggagcggt gtccgttcga tcgcagtctc
     3721 gcggtgacgg tgccgcaggc ggcccgtgct gtcgctgaag ggagcgtcga cttggaggat
     3781 ctggacgagg agcggaaggg gtggtcgggg gagcggcttc tcgccgagct ggagcggact
     3841 cggcctgcgg acgaggatct ggcggtttgc cacggtcacc tgtgcccgga caacgtgctg
     3901 ctcgaccctc gtacctgcga ggtgaccggg ctgatcgacg tggggcgggt cggccgtgcg
     3961 gaccggcact ccgatctcgc gctggtgctg cgcgagctgg cccacgagga ggacccgtgg
     4021 ttcgggccgg agtgttccgc ggcgttcctg cgggagtacg ggcgcgggtg ggatggggcg
     4081 gtatcggagg aaaagctggc gttttaccgg ctgttggacg agttcttctg actcgagtga
     4141 ccgctccgtg taaatggagg cgctcgttga tctgagcctt gccccctgac gaacggcggt
     4201 ggatggaaga tactgctctc aagtgctgaa gcggtagctt agctccccgt ttcgtgctga
     4261 tcagtctttt tcaacacgta aaaagcggag gagttttgca attttgttgg ttgtaacgat
     4321 cctccgttga ttttggcctc tttctccatg ggcgggctgg gcgtatttga agcggacccg
     4381 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     4441 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4501 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4561 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6481 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6541 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacacta aattgtaagc
     6601 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     6661 taggccgaaa tcggcaaaat cccttataaa tcaaaaga
//
