LOCUS       V004215                 6858 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004215
VERSION     V004215
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6858)
  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  JOURNAL   Unpublished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 6858)
  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 6858)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6858)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            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"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6858
                     /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
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             2296..2307
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             2320..2343
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             3293..4288
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(4558..4576)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4597..4613)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4621..4637)
                     /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(4645..4675)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4690..4711)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4999..5587)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5761..6618)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6619..6723)
                     /label="AmpR promoter"
     rep_origin      complement(join(6750..6858,1..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 ttatgacaca
     3301 agaatccctg ttacttctcg accgtattga ttcggatgat tcctacgcga gcctgcggaa
     3361 cgaccaggag ttctgggagc cgctggcccg ccgagccctg gaggagctcg ggctgccggt
     3421 gccgccggtg ctgcgggtgc ccggcgagag caccaacccc gtactggtcg gcgagcccgg
     3481 cccggtgatc aagctgttcg gcgagcactg gtgcggtccg gagagcctcg cgtcggagtc
     3541 ggaggcctac gcggtcctgg cggacgcccc ggtgccggtg ccccgcctcc tcggccgcgg
     3601 cgagctgcgg cccggcaccg gagcctggcc gtggccctac ctggtgatga gccggatgac
     3661 cggcaccacc tggcggtccg cgatggacgg cacgaccgac cggaacgcgc tgctcgccct
     3721 ggcccgcgaa ctcggccggg tgctcggccg gctgcacagg gtgccgctga ccgggaacac
     3781 cgtgctcacc ccccattccg aggtcttccc ggaactgctg cgggaacgcc gcgcggcgac
     3841 cgtcgaggac caccgcgggt ggggctacct ctcgccccgg ctgctggacc gcctggagga
     3901 ctggctgccg gacgtggaca cgctgctggc cggccgcgaa ccccggttcg tccacggcga
     3961 cctgcacggg accaacatct tcgtggacct ggccgcgacc gaggtcaccg ggatcgtcga
     4021 cttcaccgac gtgtatgcgg gagactcccg ctacagcctg gtgcaactgc atctcaacgc
     4081 cttccggggc gaccgcgaga tcctggccgc gctgctcgac ggggcgcagt ggaagcggac
     4141 cgaggacttc gcccgcgaac tgctcgcctt caccttcctg cacgacttcg aggtgttcga
     4201 ggagaccccg ctggatctct ccggcttcac cgatccggag gaactggcgc agttcctctg
     4261 ggggccgccg gacaccgccc ccggcgcctg actcgagtga ccgctccgtg taaatggagg
     4321 cgctcgttga tctgagcctt gccccctgac gaacggcggt ggatggaaga tactgctctc
     4381 aagtgctgaa gcggtagctt agctccccgt ttcgtgctga tcagtctttt tcaacacgta
     4441 aaaagcggag gagttttgca attttgttgg ttgtaacgat cctccgttga ttttggcctc
     4501 tttctccatg ggcgggctgg gcgtatttga agcggacccg gtacccagct tttgttccct
     4561 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5401 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6721 gcgcacattt ccccgaaaag tgccacacta aattgtaagc gttaatattt tgttaaaatt
     6781 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6841 cccttataaa tcaaaaga
//
