LOCUS       40924_48553        8411 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pZR606, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8411)
  AUTHORS   Chen K, Xu X, Gu L, Hildreth M, Zhou R.
  TITLE     Simultaneous gene inactivation and promoter reporting in 
            cyanobacteria
  JOURNAL   Appl. Microbiol. Biotechnol. 99 (4), 1779-1793 (2015)
  PUBMED    25434810
REFERENCE   2  (bases 1 to 8411)
  AUTHORS   Chen K, Xu X, Gu L, Zhou R.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2014) Biology and Microbiology, South Dakota State
            University, 1110 Rotunda Ln. N, Brookings, SD 57007, USA
REFERENCE   3  (bases 1 to 8411)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8411)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol."; date: "2015"; volume: "99"; issue: "4"; 
            pages: "1779-1793"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2014) Biology and Microbiology, South Dakota State 
            University, 1110 Rotunda Ln. N, Brookings, SD 57007, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(476..1894)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLEVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(1895..2340)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2749..3540
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    3849..3883
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             3910..4626
                     /codon_start=1
                     /label=Cycle 3 GFP
                     /note="Cycle 3 GFP (Crameri et al., 1996)"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             5449..6237
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(6583..6599)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7356..7944
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(8130..8270)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 gctggctgtt ttacgcgtat gacaggctcc ggaagacggt tgttgcgcac gtattcggtg
       61 aacgcactat ggcgacgctg gggcgtctta tgagcctgct gtcacccttt gacgtggtga
      121 tatggatgac ggatggctgg ccgctgtatg aatcccgcct gaagggaaag ctgcacgtaa
      181 tcagcaagcg atatacgcag cgaattgagc ggcataacct gaatctgagg cagcacctgg
      241 cacggctggg acggaagtcg ctgtcgttct caaaatcggt ggagctgcat gacaaagtca
      301 tcgggcatta tctgaacata aaacactatc aataagttgg agtcattacc aaaaggttag
      361 gaatacggtt agccatttgc ctgcttttat atagttcata tgggattcac ctttatgttg
      421 ataagaaata aaagaaaatg ccaataggat atcggcattt tcttttgcgt ttttatttgt
      481 taactgttaa ttgtccttgt tcaaggatgc tgtctttgac aacagatgtt ttcttgcctt
      541 tgatgttcag caggaagctt ggcgcaaacg ttgattgttt gtctgcgtag aatcctctgt
      601 ttgtcatata gcttgtaatc acgacattgt ttcctttcgc ttgaggtaca gcgaagtgtg
      661 agtaagtaaa ggttacatcg ttaggatcaa gatccatttt taacacaagg ccagttttgt
      721 tcagcggctt gtatgggcca gttaaagaat tagaaacata accaagcatg taaatatcgt
      781 tagacgtaat gccgtcaatc gtcatttttg atccgcggga gtcagtgaac aggtaccatt
      841 tgccgttcat tttaaagacg ttcgcgcgtt caatttcatc tgttactgtg ttagatgcaa
      901 tcagcggttt catcactttt ttcagtgtgt aatcatcgtt tagctcaatc ataccgagag
      961 cgccgtttgc taactcagcc gtgcgttttt tatcgctttg cagaagtttt tgactttctt
     1021 gacggaagaa tgatgtgctt ttgccatagt atgctttgtt aaataaagat tcttcgcctt
     1081 ggtagccatc ttcagttcca gtgtttgctt caaatactaa gtatttgtgg cctttatctt
     1141 ctacgtagtg aggatctctc agcgtatggt tgtcgcctga gctgtagttg ccttcatcga
     1201 tgaactgctg tacattttga tacgtttttc cgtcaccgtc aaagattgat ttataatcct
     1261 ctacaccgtt gatgttcaaa gagctgtctg atgctgatac gttaacttgt gcagttgtca
     1321 gtgtttgttt gccgtaatgt ttaccggaga aatcagtgta gaataaacgg atttttccgt
     1381 cagatgtaaa tgtggctgaa cctgaccatt cttgtgtttg gtcttttagg atagaatcat
     1441 ttgcatcgaa tttgtcgctg tctttaaaga cgcggccagc gtttttccag ctgtcaatag
     1501 aagtttcgcc gactttttga tagaacatgt aaatcgatgt gtcatccgca tttttaggat
     1561 ctccggctaa tgcaaagacg atgtggtagc cgtgatagtt tgcgacagtg ccgtcagcgt
     1621 tttgtaatgg ccagctgtcc caaacctcca ggccttttgc agaagagata tttttaattg
     1681 tggacgaatc gaattcagga acttgatatt tttcattttt ttgctgttca gggatttgca
     1741 gcatatcatg gcgtgtaata tgggaaatgc cgtatgtttc cttatatggc ttttggttcg
     1801 tttctttcgc aaacgcttga gttgcgcctc ctgccagcag tgcggtagta aaggttaata
     1861 ctgttgcttg ttttgcaaac tttttgatgt tcatcgttca tgtctccttt tttatgtact
     1921 gtgttagcgg tctgcttctt ccagccctcc tgtttgaaga tggcaagtta gttacgcaca
     1981 ataaaaaaag acctaaaata tgtaaggggt gacgccaaag tatacacttt gccctttaca
     2041 cattttaggt cttgcctgct ttatcagtaa caaacccgcg cgatttactt ttcgacctca
     2101 ttctattaga ctctcgtttg gattgcaact ggtctatttt cctcttttgt ttgatagaaa
     2161 atcataaaag gatttgcaga ctacgggcct aaagaactaa aaaatctatc tgtttctttt
     2221 cattctctgt attttttata gtttctgttg catgggcata aagttgcctt tttaatcaca
     2281 attcagaaaa tatcataata tctcatttca ctaaataata gtgaacggca ggtatatgtg
     2341 atgggttaaa aaggatcgat cctctagcta gagtcgacct gcatccctta acttacttat
     2401 taaataattt atagctattg aaaagagata agaattgttc aaagctaata ttgtttaaat
     2461 cgtcaattcc tgcatgtttt aaggaattgt taaattgatt ttttgtaaat attttcttgt
     2521 attctttgtt aacccatttc ataacgaaat aattatactt ttgtttatct ttgtgtgata
     2581 ttcttgattt ttttctactt aatctgataa gtgagctatt cactttaggt ttaggatgaa
     2641 aactagagga tctcaatgaa tattggttga cacgggcgta taagacatgt tatactgttg
     2701 aataacaagg acggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2761 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2821 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2881 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     2941 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3001 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3061 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3121 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3181 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3241 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3301 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3361 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3421 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3481 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3541 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3601 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3661 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccggggat
     3721 cctctagact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     3781 ccttaaacgc ctggtgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt
     3841 cgatatctag atctgcggcc gcaccggtac tagtgggccc gggtagctag ttaagaagga
     3901 gatatacata tggctagcaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
     3961 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     4021 gctacatacg gaaagcttac ccttaaattt atttgcacta ctggaaaact acctgttcca
     4081 tggccaacac ttgtcactac tttctcttat ggtgttcaat gcttttcccg ttatccggat
     4141 catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaacgc
     4201 actatatctt tcaaagatga cgggaactac aagacgcgtg ctgaagtcaa gtttgaaggt
     4261 gatacccttg ttaatcgtat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
     4321 ctcggacaca aactcgagta caactataac tcacacaatg tatacatcac ggcagacaaa
     4381 caaaagaatg gaatcaaagc taacttcaaa attcgccaca acattgaaga tggatccgtt
     4441 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
     4501 gacaaccatt acctgtcgac acaatctgcc ctttcgaaag atcccaacga aaagcgtgac
     4561 cacatggtcc ttcttgagtt tgtaactgct gctgggatta cacatggcat ggatgagctc
     4621 tacaaataat gaattaaaat aaaaaagggg acctctaggg tccccaatta attagtaata
     4681 taatctatta aaggtcattc aaaaggtcat ccaccggatc aattcccctg ctcgcgcagg
     4741 ctgggtgcca agctctcggg taacatcaag gcccgatcct tggagccctt gccctcccgc
     4801 acgatgatcg tgccgtgatc gaaatccaga tccttgaccc gcagttgcaa accctcactg
     4861 atccgcatgc ccgttccata cagaagctgg gcgaacaaac gatgctcgcc ttccagaaaa
     4921 ccgaggatgc gaaccacttc atccggggtc agcaccaccg gcaagcgccg cgacggccga
     4981 ggtcttccga tctcctgaag ccagggcaga tccgtgcaca gcaccttgcc gtagaagaac
     5041 agcaaggccg ccaatgcctg acgatgcgtg gagaccgaaa ccttgcgctc gttcgccagc
     5101 caggacagaa atgcctcgac ttcgctgctg cccaaggttg ccgggtgacg cacaccgtgg
     5161 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     5221 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     5281 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     5341 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     5401 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacatcat gagggaagcg
     5461 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     5521 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     5581 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     5641 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     5701 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     5761 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     5821 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     5881 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     5941 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     6001 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     6061 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     6121 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     6181 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     6241 tgtctaacaa ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga
     6301 tgcactaagc acataattgc tcacagccaa actatcaggt caagtctgct tttattattt
     6361 ttaagcgtgc ataataagcc ctacacaaat tgggagatat atcatgaaag gctggctttt
     6421 tcttgttatc gcaatagttg gcgaagtaat cgcaacatcc gcattaaaat ctagcgaggg
     6481 ctttactaag ctgatccggt ggatgacctt ttgaatgacc tttaatagat tatattacta
     6541 attaattggg gaccctagag gtcccctttt ttattttaat tcactggccg tcgttttaca
     6601 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     6661 cttcgccagg ggggatccac taggccttaa gatcgaaaga gctggtagtt ggcgcactgt
     6721 tcgaagaact gccgatgtcc agtaagattc ttactatgct ggttgaaccg gatgctggta
     6781 aagctacttg ggttgctgct tctacttatg gtaccgatac aactactggt gaggaagtta
     6841 aaggagctcg cgaaagcttg catgcctgca gcaatggcaa caacgttgcg caaactatta
     6901 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6961 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7021 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7081 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7141 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7201 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7261 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7321 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7381 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7441 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7501 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7561 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7621 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7681 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7741 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7801 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7861 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7921 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7981 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8041 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8101 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     8161 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     8221 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     8281 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     8341 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     8401 gcgcgaggca g
//
