LOCUS       sequence-232096-        7670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RpoD5 epitope tagged construct for integration at neutral site 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7670)
  AUTHORS   Fleming KE, O'Shea EK
  TITLE     An RpaA-Dependent Sigma Factor Cascade Sets the Timing of Circadian 
            Transcriptional Rhythms in Synechococcus elongatus.
  JOURNAL   Cell Rep. 2018 Dec 11;25(11):2937-2945.e3. doi: 
            10.1016/j.celrep.2018.11.049.
  PUBMED    30540929
REFERENCE   2  (bases 1 to 7670)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2018.11"; journalName: "Cell Rep"; date: 
            "2018-12-11- 11"; volume: "25"; issue: "11"; pages: "2937-2945.e3"
COMMENT     Sequence Label: KFP13
FEATURES             Location/Qualifiers
     source          1..7670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     202..224
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             2977..3042
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3927..4718
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6872..7460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7361..7380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7614..7631
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(7646..7670,1..116)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
      121 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
      181 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
      241 cgaaacgcgc gaggcaggat cgatccggca gccggcggag cgctgctttc ttggcaagcg
      301 gtcgccagcc ccaacgccag ggctgccagc ccgaaacagc ggggcaaggc agcttggaag
      361 ggcgatcgca gcacgggcat ggcaatgtct ctctgaagga atgcagacct tattcgtaca
      421 gccagggttg aatcgtgggg gtccaatcac ttagctctgc tgggctaaac cagagagcaa
      481 tttcctgttg tgctgtttcg attgcatccg agccatggat gatgttgcgg ccaatattga
      541 caccaaaatc accacggatg gtgcccggtt ctgccgtcag cggattggta gcgccgatca
      601 acttgcgagc agccgccaca acgccttcgc cttccaagac gatcgccacg atcggcccag
      661 aggtgatgaa ctcgacgagg ccattgaaga aggggcgctc gcggtggaca gcatagtgct
      721 gttcggccag ctcgcgactg ggcttcagct gctttaggcc caccagtttg aagccttttt
      781 gctcaaagcg gccgatgatc gtaccgacca aaccccgctg aacgccatcg ggcttgatgg
      841 caataaatgt gcgttccaca gacatctaga tagtcctcaa gacgaggcaa gcattgagct
      901 tgccttccta tggttcggga tcactgggat tcttgacaag cgatcgcggt cacatcgcta
      961 tctcttagga cttcgcagcg ggcgagtcgg attgacccgg tagggatttc gccagatcaa
     1021 tgcccgtggt ttgtttcagc ttctccagca agctagcgat ttgggtagcg ctgccttccc
     1081 cttcgccaat cacagtgatc gactccacgt cgatatctgg cacggtgcct gaaagcgtga
     1141 cgagcaggga ctcgaagctt gcatgcctgc aggaaatgaa gagcatgaag cgctaaagag
     1201 ttccgaccca ttcctgggaa ttgcggggaa cctccatcat actcggcgct gtcgagaaac
     1261 aagaatacag ccaagcgggg ctggacgccg cattgagaaa gatttttgca gctctaacag
     1321 ccccaagcca gcaggcaagc accctttcca gcgcccccag agcccagagg ctcactcata
     1381 gaatcacgat tctcctaggg caaagccgca atggccctct aggctggatt ccgagtaggg
     1441 gagtcatttc agacgagcga tcgccaacaa tgcaaactca gcgttgatgc cacatgtttt
     1501 ctcagaattc ttgatggttc tcaatggtga gctagaggga tacaaaagtt caagaaacca
     1561 agcactgaac tgggcaaata tcaaagcaac atctgggctt taccgaggac ttacattttg
     1621 gccaaaatct aaagagagct ggggcactgg caagaatttt attttttata gagagactat
     1681 tttccagata agcactggcc ttgaagtcga cggcaaaata ttgcatcgct ccagccagtc
     1741 gttatccctg taataaactg gatattcacc atgacggctg cctatttcac tcattccacg
     1801 gaatacgaag cagtttcgtc tgacgtcgct aacttgtcag aaactccaag ttctgaagcc
     1861 gtggaaacag cggtctctag ccgccagtca accgatctga ttcggctgta tttacaagac
     1921 atcgggcgag tccgtctcct tgggcgcgat gaagaggtga gtgaggcacg gtgggtgcag
     1981 cgctacatgc agttgttggc ggttcggaat tgcgtcgccg ttgaagaagg aggggttctc
     2041 aaggattacg ttgacctgca ggaaggccgc gatcgcctga ccgcgcagct gggtcaccgt
     2101 ccttcctttg aagcttgggc caagattgtc gattgttcct tgactgaatt acgccaagca
     2161 ctacaactgg ggctgcatcg ctgggctgaa gtctgccagc tcgatcttga ggaattaaag
     2221 caggtctgca aagaaggcca gcgagctaaa gatcacatga tcaaggccaa cctccgttta
     2281 gttgtttccg tcgctaaaaa atatcaaaac cgaggactag agcttctcga tctcgttcaa
     2341 gaaggcaccc taggactaga gcgggctgtt gagaagttcg acccgacgaa aggctatcga
     2401 tttagtacct atgcctattg gtggattcgt cagggaatta cccgcgcgat cgcaacccaa
     2461 agtcggacga ttcgcttgcc cgtccatgtg acggagaaat taaaccggat caaaaaagcc
     2521 cagcgcaaaa ttgcgcaaac aaagggcaga actgccaacc ttgaggagat tgctcaagaa
     2581 cttgagatga cccccgatca agtgcgggaa gtcctgttgc gggtgcccaa atcgatttcc
     2641 ctagaaaccc gtgttggcaa ggaaaaggat acggagctag gggatttgct ggaaaccacc
     2701 gatttcaccc cagaagaacg attgactcag gaatccctca gcaatgatct gcagcggtta
     2761 ttgctcgaac tcaatcaccg cgaacgcgat gtaattcgga tgcggtttgg tctgcgcgat
     2821 gggcagattt attccctcgc tgaaattggg caagtgctcg acctatcgcg tgagcgagtt
     2881 cgccagattg aggctaaggc cctgcaaaag ctgcggcatc cgcgacgacg caatcagatc
     2941 cgtgactatt tggagttgtt gcacggcagc ggcagcgatt acaaagatca cgatggcgat
     3001 tacaaagatc acgatatcga ttacaaagat gatgatgata aatagacccg ggctcgagct
     3061 agcaagcttc tctagagctt tatgcttgta aaccgttttg tgaaaaaatt tttaaaataa
     3121 aaaaggggac ctctagggtc cccaattaat tagtaatata atctattaaa ggtcattcaa
     3181 aaggtcatcc accggatcaa ttcccctgct cgcgcaggct gggtgccaag ctctcgggta
     3241 acatcaaggc ccgatccttg gagcccttgc cctcccgcac gatgatcgtg ccgtgatcga
     3301 aatccagatc cttgacccgc agttgcaaac cctcactgat ccgcatgccc gttccataca
     3361 gaagctgggc gaacaaacga tgctcgcctt ccagaaaacc gaggatgcga accacttcat
     3421 ccggggtcag caccaccggc aagcgccgcg acggccgagg tcttccgatc tcctgaagcc
     3481 agggcagatc cgtgcacagc accttgccgt agaagaacag caaggccgcc aatgcctgac
     3541 gatgcgtgga gaccgaaacc ttgcgctcgt tcgccagcca ggacagaaat gcctcgactt
     3601 cgctgctgcc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3661 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     3721 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3781 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     3841 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     3901 gtcgccctaa aacaaagtta aacatcatga gggaagcggt gatcgccgaa gtatcgactc
     3961 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     4021 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     4081 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     4141 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     4201 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     4261 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     4321 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     4381 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     4441 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     4501 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     4561 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     4621 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     4681 acgtgaaagg cgagatcacc aaggtagtcg gcaaataatg tctaacaatt cgttcaagcc
     4741 gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg cactaagcac ataattgctc
     4801 acagccaaac tatcaggtca agtctgcttt tattattttt aagcgtgcat aataagccct
     4861 acacaaattg ggagatatat catgaaaggc tggctttttc ttgttatcgc aatagttggc
     4921 gaagtaatcg caacatccgc attaaaatct agcgagggct ttactaagct gatccggtgg
     4981 atgacctttt gaatgacctt taatagatta tattactaat taattgggga ccctagaggt
     5041 cccctttttt attttaaaaa ttttttcaca aaacggttta caagcatact agaggatcgg
     5101 cggccgcgga tccccgggta ccgagctcga atttcgagct tctggagcag gaagatgtcg
     5161 cgggcattag caccagcggt ctgccaagcc tccgccagcc gttgggtccc ttccgcttga
     5221 gcttttccat cttcgacgat acgggcggcg gccccccgcg cttccgcgat cgcccgttta
     5281 caagctgcct cagctggggc gatcacatcg gcttgaagtt gctgctgcac ctgtttgatc
     5341 cgctcctgct gcacagggag ttctgcttgg ctacgagcga cttcggtagc aatgtccgct
     5401 tcagcttcgg ccaccaccgc ttcgcgccgc gtcaacgcat cctgaatccg gcgctcggcc
     5461 tcggcttggg cgatcgctac atcgcgatcg atccgacgca gggccgtgat cttgtcattt
     5521 tcggccgttt ggatcgcaga ggcagcctgg gcatcggctt cagcaattcg ggcatctcgc
     5581 tgcagatcag cccgctgctt gcgtccacta gccgagagat aaccgacctc atcggaaatg
     5641 ttctggactt gcagcgtatc gaggactaga cccagctgct caaggtcatc ctccgcctct
     5701 tccagcagac ttttggcaaa ggcaattttg tcctcgttga tctgctccgg cgtgaggctg
     5761 gctaaaacac cacgcaagtt gccttcgagg gtctccttgg caatttgctc gatttcctta
     5821 cggtttttgc caagcagccg ctcgatcgcg ttgtggatgg tcggttcttc cccagcaatc
     5881 ttgatattgg caacgccttc aacagtcagg ggaatgccgc ccttggagaa ggcattggaa
     5941 acgcgcaact caatgatcat gttggtcaga tccatgcgga gcgctttttc cagcagaggt
     6001 acccgcaggc tgctgccgcc cttgaccaag cgatagccaa ctcggcggcc atcactactg
     6061 cggcgactac tgccagcaaa gatcaaaatt tcactgggtt ggcagatgta gtagagattg
     6121 cgcaggacta agctgccagc cccggcgact caccagtcac agaaaagcat cttacggatg
     6181 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6241 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6301 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6361 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     6421 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6481 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6541 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6601 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6661 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6721 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6781 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6841 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6901 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6961 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7021 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7081 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7141 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7201 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7261 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7321 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7381 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7441 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7501 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7561 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7621 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
//
