LOCUS       62056_8645        7971 bp DNA     circular SYN 22-DEC-2019
DEFINITION  Expression vector pDU1, complete sequence.
ACCESSION   MK948095
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  AUTHORS   Niu T-C., Zhang J-Y., Zhang C-C.
  TITLE     Tight Control of Gene Expression in Anabaena using a hybrid 
            regulatory system consisting of a copper inducible promoter and a 
            theophylline riboswitch
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7971)
  AUTHORS   Niu T-C., Zhang J-Y., Zhang C-C.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-MAY-2019) Center for Algal Biology and Applied 
            Research, Institute of Hydrobiology, Donghu Nanlu, Wuhan, Hubei 
            430072, 086
REFERENCE   3  (bases 1 to 7971)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-MAY-2019) Center for Algal Biology and Applied Research, 
            Institute of Hydrobiology, Donghu Nanlu, Wuhan, Hubei 430072, 086"
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3801..7971
                     /mol_type="other DNA"
                     /db_xref="taxon:28072"
                     /organism="Nostoc sp. PCC 7524"
     CDS             596..1387
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    1943..2083
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2269..2857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      2952..2976
                     /label=terminator
                     /note="terminator"
                     /regulatory_class="terminator"
     regulatory      2983..3448
                     /note="petE; the promoter region(-475 to -9) of Anabaena
                     petE (id:all0258) gene"
                     /regulatory_class="promoter"
     regulatory      3455..3517
                     /label=theophylline riboswitch
                     /note="theophylline riboswitch"
                     /regulatory_class="ribosome_binding_site"
     misc_feature    3530..3568
                     /label=protein domain linker
                     /note="protein domain linker"
     CDS             3563..3652
                     /label=Twin-Strep-tag
                     /note="two Strep-Tag(R)II moieties connected by a linker 
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
     CDS             3656..3673
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      3740..3787
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     gene            4878..5999
                     /gene="repA"
                     /label=repA
     CDS             4878..5999
                     /codon_start=1
                     /transl_table=11
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="required for plasmid replication in cyanobacteria"
                     /protein_id="QGX42291.1"
                     /translation="MISPESLNTNVSLTLLTRELQTEIILGTYTVRVHTRIGREPCARL
                     WYLCRALDKDGSGHLTLPLPVVQTFLDCSDKSVYRWLQDGKKIGAFRRYKIKAGMITVY
                     LGGMFQVCYNLNLKRWGDVAVVPLVQVLSDLRSLTTGIVTQSFQQKSRYAANRQLKPEY
                     RKLFGAPHPNELVKDTRQSSLKSPEGEVPCVLHISSSRIFVSKSFIHYGTSQKAVSCEL
                     GIHKRTVRRHQKQLGMNRRQLCQAKIEYNQLRHARNNDASEFWAFTGTKTDIGYQVMGD
                     AVVFSDGIAPGAKKRQPNTYQIDATEFDGRLFKVGDKVFMNRCNIYREQFTLTTMSAAR
                     RKYHFKLSQCHFSENRAGRVGNRFVIGCHSGEI"
     CDS             complement(6620..7171)
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="required for plasmid replication in cyanobacteria"
                     /protein_id="QGX42292.1"
                     /translation="MKVNGNGRAKILTSDELRRLFSDGFTTPRDRVLFGICLFTGCRVS
                     EALALQTTDIKGETLTFRKSTTKGKLKTRVVDIQPGLAALMADYHPKPGTLFPGMRGVS
                     DRLTRYAADKILRDAAKRIGLEGISTHSFRRTALNQMSSAGIPLRHIQEISGHNDLGTL
                     QRYLEVTPEQRRKAVSVIGF"
     CDS             complement(7543..7896)
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="required for plasmid replication in cyanobacteria"
                     /protein_id="QGX42293.1"
                     /translation="MADKTLATFRIDSEEWESFKNLASSESSNASALLTEFVRWYLAGN
                     RFNTPTSHTPTHLDTSLEQRIDNIEQRLDKVTTNNLDNIDEFIDKRIEDNLATRLDKLQ
                     SQLEELRGKSKAR"
ORIGIN
        1 atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagagtc tagctagact
       61 ctagcccaag ccttgcgcgt catcgacatt tatacccatc gctggcggat cgaggagttc
      121 cataaggcat ggaaaaccgg agcaggagcc gagaggcaac gcatggagga gccggataat
      181 ctggagcgga tggtctcgat cctctcgttt gttgcggtca ggctgttaca gctcagagaa
      241 agcttcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta
      301 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
      361 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
      421 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
      481 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
      541 ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat
      601 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
      661 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
      721 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
      781 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
      841 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
      901 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
      961 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     1021 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     1081 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     1141 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     1201 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     1261 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     1321 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     1381 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     1441 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     1501 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1561 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     1621 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
     1681 aggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
     1741 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca
     1801 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     1861 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     1921 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     1981 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     2041 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     2101 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2161 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2221 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2281 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2341 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2401 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2461 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2521 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2581 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2641 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2701 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2761 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2821 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2881 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2941 aaaaaggatc tccccgcttc ggcggggttt ttttttggat cctaaagcct gtgaaattaa
     3001 ctgagtgctg agtactgagt actgggtgct gagactaatg ggagtggtaa gtttttaaac
     3061 tggcattcct cactcccaga caacacagga ctcagaacac agtactcaga attttttgct
     3121 gaggtactga gtacacagct aataaaattg ggcaatctcc gcgcctctat gacttgaagg
     3181 agagtgtagg ggtatagggg aaagatatct tttatctaca tcacataaat aaaaaattta
     3241 atttgtcgct ctggctgcat atattgatgt atttttagcc ataagttttt tagtgccatg
     3301 taattatagt gatttttagc gatcgcagag catttttccc tggatttatc gcgatctcaa
     3361 aaaaaatttg cccgaagtat gacagattgt catatttggt gtcgatttta tttaaaatga
     3421 aataagaaaa ataaaactac aggttaggga attcggtgat accagcatcg tcttgatgcc
     3481 cttggcagca ccctgctaag gaggtaacaa caagatgctc gagagtcccg ggggttctgg
     3541 tggtggtagc actagcgtcg gtagcgcttg gagccatccc caatttgaaa aaggtggtgg
     3601 cagcggcggt ggtagcggtg gtagcgcttg gtctcaccca caattcgaaa agtgacacca
     3661 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     3721 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3781 ttttttgctg aaaggaggaa ctatatccgg cggccgcaag cccaagcgtt ttgttattgg
     3841 gctgactcaa gatgataatg tcaagacctt aggtatcgag ggtgaaggcg gtgtccgtaa
     3901 gtgttttagg atgttacgcc tgggtaagtt tgcagttagc cacgccaagt ccttaaaaga
     3961 ccttgccctt gtggagtggt taaagtcggg gaaatatcgc tgcatggtgg atgatatgcc
     4021 gtgtcagtta ccagatgtaa gcagttacaa aatggttatg ccccggttac agtctgtaac
     4081 cagttttcag ccactccaaa cggctcaacc gacctcagaa caagcattac agcccgtttc
     4141 atccctccca gaaccaagag atgaaaccct agcaaaagtg gtaaaagcct gtctggaagc
     4201 gggtttttca gaaagtaagg tgattaaaga aattttgggc tatcaaggcg ggcgttacca
     4261 agaaggtaaa tcgctactgg aaaaacttaa ggatttgtag cggtgcggtt tagcattgac
     4321 tgttacttgc tgtacaagga taattttctc aaagcggaat ggtgttaaac ccgattatta
     4381 gagtcgtcat agcaaaagct tgaaaactgg caagggtgta ctggttttcg ttctttttgg
     4441 ttctaatggt tattttttac cagtttattt agtaataacc gagtgccaag tttcttgttt
     4501 tacatagcaa agttagcaaa aaacattttg ccgtgaattt ccatctgtaa aaagcgtcac
     4561 ggcgaaattc agtatttagt aaatacaaaa ataagtcaaa cgttgatcat gcaacacttt
     4621 ggcgtacaat tgattgaaaa ttttgccgat aggcaaagtt taaattacct attaaaaaaa
     4681 gttcagtgct tgggggttgt tatgctactt aaccgaagct aaataaaaca cacatacaca
     4741 gaattgttac cacaccaaac accaaacaga aaaaacgtgc tttgaattta tcaattaatg
     4801 ggtgtagtta acaagtttta tttagcttca ggttaagcaa ataacattca ataggtactg
     4861 gactgagaca tttccaaatg atcagcccag aaagcttaaa tacaaatgta tcactaaccc
     4921 tgcttacaag ggaactacaa actgaaataa ttcttggcac ttacacggtc agggtacaca
     4981 ctcgcattgg tcgagaacct tgtgcgcggt tgtggtattt gtgtcgagcg cttgataagg
     5041 atgggtctgg tcatttaaca ttgccattgc ctgtagtaca gacgttctta gattgtagtg
     5101 ataagtctgt ttaccgttgg ttacaggatg gaaaaaaaat aggagcattc cgccgctaca
     5161 aaattaaagc aggaatgata accgtctact tgggcggtat gtttcaagtt tgttacaacc
     5221 taaatttaaa aagatggggt gatgttgcag tagtgccttt agtccaagtg ttaagcgatt
     5281 tacgctcact gaccactggc atagtcacac aaagctttca acaaaagagc cgctatgcgg
     5341 ccaacagaca acttaaacca gagtacagaa aactttttgg agcgccacac ccgaacgagc
     5401 tagtgaagga cacaaggcaa tcctctctta aatcgccaga gggggaagta ccttgtgtac
     5461 ttcacattag ttcgtcgcgg atattcgtct caaaaagttt catccactat ggtacatctc
     5521 aaaaagctgt cagctgtgag ttgggtattc acaagcgcac agtgcgacga catcaaaagc
     5581 agctgggcat gaatcgccgt caactttgcc aagccaaaat tgagtacaat caactccggc
     5641 acgctcggaa taatgacgcg tctgaatttt gggctttcac tggaaccaaa actgatatcg
     5701 gttatcaagt tatgggtgac gcggttgtgt tttctgatgg cattgcccct ggagccaaaa
     5761 agagacagcc caacacttac caaattgacg cgactgaatt tgacggtcga ttattcaaag
     5821 tgggtgacaa ggtgttcatg aatcggtgca acatctaccg cgaacaattc accctaacaa
     5881 cgatgtcagc agcacggcgg aagtaccatt tcaaactgtc acagtgccac ttcagcgaaa
     5941 atcgagccgg tcgggtaggc aaccgttttg taataggttg ccactctggc gagatatgaa
     6001 ggacattttt aagaattttt ttttagcgtg aaatgctgcg gcaaccagta tagtgatcga
     6061 cacgaactag tgcagcatga ccgagagcct atagggtcgc ggctttttgg caatagagat
     6121 cgtatgcaga tcatgcgtag tactttcttc gaccttaacc ccaaagaaag cttattagat
     6181 ggagaaggtc acaattagaa aatattcatc gtcccgattg acccttaacc tttaatcggt
     6241 ttccacttgt aaaaaattcg gcgagccatt cttcttaacc ctcagagtcg aacttcatac
     6301 aagaatatac aaaccctttg ggtttaccac taaaacaacg gcgataaaaa cctaataggt
     6361 ttccacagaa acgtctaagg atttaggcgt gttgttttgg ggcaatttga gaaaatatta
     6421 aaccagtcac aagttaactt gtcgaagaaa tattgcagca ttctgcaaca cagggaaagg
     6481 tataacgttt ccaaagtccg gaacgtgtgc agagttctag aaccgatgga tactttaaaa
     6541 gtatcctcaa gtgtagagcg tgtgtttagg ctgcgtcttt ttttagcacc cataggggtc
     6601 taaacagcgt tggcgtacat tagaagccaa tcacggatac agctttgcgt cgctgttcgg
     6661 gtgtaacttc aagatagcgt tgcagtgtgc caaggtcatt gtgaccggat atctcttgaa
     6721 tgtgtcgcaa cgggataccg gcgctagaca tttggttgag ggcagtacgg cggaaactgt
     6781 gggtactgat gccttctagc ccgattcttt tggctgcatc gcgcaagatt ttatccgccg
     6841 cgtatcgcgt gagcctatcg ctgactcccc tcatgccagg gaacagggtt cccggtttgg
     6901 ggtgatagtc agccatgagt gcggctagtc ctggctggat gtcaaccacg cgggttttga
     6961 gtttcccttt ggtggtagac ttcctaaagg ttagtgtttc gcctttaatg tccgttgttt
     7021 ggagtgctag agcttcacta acgcggcaac cggtgaatag acagatgcca aacaaaacgc
     7081 gatcgcgcgg tgtggtgaat ccgtcgctaa acagtcgcct gagttcgtcg gaggtgagta
     7141 ttttggctcg tccgttgccg ttaaccttca taacccttac agattaagca ttccaatact
     7201 ctacacgtta tctgcatttt gtggaattgg tggctatatc tattgattct tcgtatgctt
     7261 tctgttgaaa gccagtgcta gtttagccaa gcttatcaat cgttgcacgg taaaatcagg
     7321 ctttcgacgt aaggacagaa ttttgataac ttcagcttat gttgtctcct gaagaaagca
     7381 actctttgat aaacgcgtcg atggctttcc cggcggctga ctgcctaccc atttttagct
     7441 tattgagggt gcgatcgcgg gttgcctcca atttgtctag attgtctata ctattgtcta
     7501 tattctttct gtcttgcccg gtagcttgtc cttctgcctg aactaccggg ctttcgattt
     7561 tccccgcagt tcctccagtt gcgattgaag tttgtctaga cgtgttgcta gattatcttc
     7621 tatacgcttg tctataaatt catctatatt gtctagatta ttagttgtga ctttatctag
     7681 acgttgttca atattgtcta tacgctgttc gagggatgtg tctagatggg tgggagtgtg
     7741 agaagtggga gtattaaacc tgttacctgc caaataccaa cgaacaaatt ctgttaacag
     7801 tgctgaggcg ttggaacttt cagaactagc aaggttttta aaagactccc attcttcgga
     7861 gtcaatacga aaagtggcta gagttttgtc tgccatgatt agataatgtc tatacttgta
     7921 tagatagcag tataaactat tgtctatatt agtgtctata taagtctaga t
//
