LOCUS       40924_33832        6497 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pOpt_gLuc_Paro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  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  JOURNAL   Unpublished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 6497)
  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 6497)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6497)
  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            "Unpublished"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..6497
                     /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             2095..2106
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             2119..2142
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     promoter        complement(4197..4215)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4236..4252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4260..4276)
                     /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(4284..4314)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4329..4350)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4638..5226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5400..6257)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6258..6362)
                     /label=AmpR promoter
     rep_origin      complement(join(6389..6497,1..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 agggcaggaa
     1261 gccgacggag aacaacgagg acttcaacat tgtcgccgtg gcgtccaact ttgccaccac
     1321 cgatctggac gccgaccgcg gcaagctgcc gggcaagaag ctgcccctgg aggtcctcaa
     1381 ggaaatggag gcgaacgccc ggaaggccgg ctgcacccgc ggctgcctga tctgcctgtc
     1441 ccacatcaag tgcacgccga agatgaagaa gttcattccc ggccggtgag cttgcggggt
     1501 tgcgagcaac actccagcaa cgaacagtgc ccaagtcagg aatctgcagt cagcctgggc
     1561 tttcggcggc tttttcttgg gcaaacagct tgcactcatg ccagcgcggc ttgtccagcc
     1621 tcacttgagc tttccagctg ctaccagccg ggctatacga cagcgacaga gccatagcgt
     1681 ggaatcactt atttgggttg ccgaagtagc ggtcggagcg tgagttcttg gtcaagccgc
     1741 cccttatccg gttcctgtcc gtgtctttgt ccctcgttca cccttcgcgg cacccttcat
     1801 ccccttgctt gcaggtgcca cacctacgag ggcgacaagg agtccgcgca gggcggcatt
     1861 ggcgaggcga tcgtggacat ccccgagatt ccgggcttca aggacctgga gccgatggag
     1921 cagttcatcg cccaggtgga cctgtgcgtg gactgcacga ccggctgtct gaagggcctg
     1981 gccaatgtgc agtgttccga cctgctcaag aagtggctgc cccagcgctg tgccaccttc
     2041 gccagcaaga tccagggcca ggtggacaag attaagggcg ctggcggcga tacgatcgag
     2101 ggcagggata tcgaattctg gagccacccg cagttcgaga agtaaccgct ccgtgtaaat
     2161 ggaggcgctc gttgatctga gccttgcccc ctgacgaacg gcggtggatg gaagatactg
     2221 ctctcaagtg ctgaagcggt agcttagctc cccgtttcgt gctgatcagt ctttttcaac
     2281 acgtaaaaag cggaggagtt ttgcaatttt gttggttgta acgatcctcc gttgattttg
     2341 gcctctttct ccatgggcgg gctgggcgta tttgaagcgg gatccttcga acgtacggtt
     2401 aactcgcgac ctgcaggtac gtaatttaaa tacgcgtgct gaggcttgac atgattggtg
     2461 cgtatgtttg tatgaagcta caggactgat ttggcgggct atgagggcgg gggaagctct
     2521 ggaagggccg cgatggggcg cgcggcgtcc agaaggcgcc atacggcccg ctggcggcac
     2581 ccatccggta taaaagcccg cgaccccgaa cggtgacctc cactttcagc gacaaacgag
     2641 cacttatagg tacccgacta ttctgccgct atacataacc actcagctag cttaagatcc
     2701 catccctagg gcatgccggg cgcgccagaa ggagcgcagc caaaccagga tgatgtttga
     2761 tggggtattt gagcacttgc aacccttatc cggaagcccc ctggcccaca aaggctaggc
     2821 gccaatgcaa gcagttcgca tgcagcccct ggagcggtgc cctcctgata aaccggccag
     2881 ggggcctatg ttctttactt ttttacaaga gaagtcactc aacatcttaa aatggccagg
     2941 tgagtcgacg agcaagcccg gcggatcagg cagcgtgctt gcagatttga cttgcaacgc
     3001 ccgcattgtg tcgacgaagg cttttggctc ctctgtcgct gtctcaagca gcatctaacc
     3061 ctgcgtcgcc gtttccattt gcaggaagct tactccgccc tccccggtgc tgaagaattt
     3121 cgaagcatgg acgatgcgtt gcgtgcactg cggggtcggt atcccggttg tgagtgggtt
     3181 gttgtggagg atggggcctc gggggctggt gtttatcggc ttcggggtgg tgggcgggag
     3241 ttgtttgtca aggtggcagc tctgggggcc ggggtgggct tgttgggtga ggctgagcgg
     3301 ctggtgtggt tggcggaggt ggggattccc gtacctcgtg ttgtggaggg tggtggggac
     3361 gagagggtcg cctggttggt caccgaagcg gttccggggc gtccggccag tgcgcggtgg
     3421 ccgcgggagc agcggctgga cgtggcggtg gcgctcgcgg ggctcgctcg ttcgctgcac
     3481 gcgctggact gggagcggtg tccgttcgat cgcagtctcg cggtgacggt gccgcaggcg
     3541 gcccgtgctg tcgctgaagg gagcgtcgac ttggaggatc tggacgagga gcggaagggg
     3601 tggtcggggg agcggcttct cgccgagctg gagcggactc ggcctgcgga cgaggatctg
     3661 gcggtttgcc acggtcacct gtgcccggac aacgtgctgc tcgaccctcg tacctgcgag
     3721 gtgaccgggc tgatcgacgt ggggcgggtc ggccgtgcgg accggcactc cgatctcgcg
     3781 ctggtgctgc gcgagctggc ccacgaggag gacccgtggt tcgggccgga gtgttccgcg
     3841 gcgttcctgc gggagtacgg gcgcgggtgg gatggggcgg tatcggagga aaagctggcg
     3901 ttttaccggc tgttggacga gttcttctga ctcgagtgac cgctccgtgt aaatggaggc
     3961 gctcgttgat ctgagccttg ccccctgacg aacggcggtg gatggaagat actgctctca
     4021 agtgctgaag cggtagctta gctccccgtt tcgtgctgat cagtcttttt caacacgtaa
     4081 aaagcggagg agttttgcaa ttttgttggt tgtaacgatc ctccgttgat tttggcctct
     4141 ttctccatgg gcgggctggg cgtatttgaa gcggacccgg tacccagctt ttgttccctt
     4201 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4261 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4321 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4441 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4501 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4561 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4801 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5041 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5221 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5341 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5401 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5461 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5521 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5581 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5641 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5701 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5761 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5821 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5881 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5941 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6001 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6061 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6121 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6181 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6241 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6361 cgcacatttc cccgaaaagt gccacactaa attgtaagcg ttaatatttt gttaaaattc
     6421 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     6481 ccttataaat caaaaga
//
