LOCUS       sequence-226890-        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"
     rep_origin      725..1313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1214..1233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1467..1484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1499..1639
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1565..1584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1725..1747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             4500..4565
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5450..6241
                     /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"
ORIGIN
        1 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
       61 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      121 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      181 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
      241 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      301 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      361 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      421 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      481 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      541 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      841 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
      901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1381 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1441 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1501 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     1561 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     1621 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1741 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag gatcgatccg gcagccggcg
     1801 gagcgctgct ttcttggcaa gcggtcgcca gccccaacgc cagggctgcc agcccgaaac
     1861 agcggggcaa ggcagcttgg aagggcgatc gcagcacggg catggcaatg tctctctgaa
     1921 ggaatgcaga ccttattcgt acagccaggg ttgaatcgtg ggggtccaat cacttagctc
     1981 tgctgggcta aaccagagag caatttcctg ttgtgctgtt tcgattgcat ccgagccatg
     2041 gatgatgttg cggccaatat tgacaccaaa atcaccacgg atggtgcccg gttctgccgt
     2101 cagcggattg gtagcgccga tcaacttgcg agcagccgcc acaacgcctt cgccttccaa
     2161 gacgatcgcc acgatcggcc cagaggtgat gaactcgacg aggccattga agaaggggcg
     2221 ctcgcggtgg acagcatagt gctgttcggc cagctcgcga ctgggcttca gctgctttag
     2281 gcccaccagt ttgaagcctt tttgctcaaa gcggccgatg atcgtaccga ccaaaccccg
     2341 ctgaacgcca tcgggcttga tggcaataaa tgtgcgttcc acagacatct agatagtcct
     2401 caagacgagg caagcattga gcttgccttc ctatggttcg ggatcactgg gattcttgac
     2461 aagcgatcgc ggtcacatcg ctatctctta ggacttcgca gcgggcgagt cggattgacc
     2521 cggtagggat ttcgccagat caatgcccgt ggtttgtttc agcttctcca gcaagctagc
     2581 gatttgggta gcgctgcctt ccccttcgcc aatcacagtg atcgactcca cgtcgatatc
     2641 tggcacggtg cctgaaagcg tgacgagcag ggactcgaag cttgcatgcc tgcaggaaat
     2701 gaagagcatg aagcgctaaa gagttccgac ccattcctgg gaattgcggg gaacctccat
     2761 catactcggc gctgtcgaga aacaagaata cagccaagcg gggctggacg ccgcattgag
     2821 aaagattttt gcagctctaa cagccccaag ccagcaggca agcacccttt ccagcgcccc
     2881 cagagcccag aggctcactc atagaatcac gattctccta gggcaaagcc gcaatggccc
     2941 tctaggctgg attccgagta ggggagtcat ttcagacgag cgatcgccaa caatgcaaac
     3001 tcagcgttga tgccacatgt tttctcagaa ttcttgatgg ttctcaatgg tgagctagag
     3061 ggatacaaaa gttcaagaaa ccaagcactg aactgggcaa atatcaaagc aacatctggg
     3121 ctttaccgag gacttacatt ttggccaaaa tctaaagaga gctggggcac tggcaagaat
     3181 tttatttttt atagagagac tattttccag ataagcactg gccttgaagt cgacggcaaa
     3241 atattgcatc gctccagcca gtcgttatcc ctgtaataaa ctggatattc accatgacgg
     3301 ctgcctattt cactcattcc acggaatacg aagcagtttc gtctgacgtc gctaacttgt
     3361 cagaaactcc aagttctgaa gccgtggaaa cagcggtctc tagccgccag tcaaccgatc
     3421 tgattcggct gtatttacaa gacatcgggc gagtccgtct ccttgggcgc gatgaagagg
     3481 tgagtgaggc acggtgggtg cagcgctaca tgcagttgtt ggcggttcgg aattgcgtcg
     3541 ccgttgaaga aggaggggtt ctcaaggatt acgttgacct gcaggaaggc cgcgatcgcc
     3601 tgaccgcgca gctgggtcac cgtccttcct ttgaagcttg ggccaagatt gtcgattgtt
     3661 ccttgactga attacgccaa gcactacaac tggggctgca tcgctgggct gaagtctgcc
     3721 agctcgatct tgaggaatta aagcaggtct gcaaagaagg ccagcgagct aaagatcaca
     3781 tgatcaaggc caacctccgt ttagttgttt ccgtcgctaa aaaatatcaa aaccgaggac
     3841 tagagcttct cgatctcgtt caagaaggca ccctaggact agagcgggct gttgagaagt
     3901 tcgacccgac gaaaggctat cgatttagta cctatgccta ttggtggatt cgtcagggaa
     3961 ttacccgcgc gatcgcaacc caaagtcgga cgattcgctt gcccgtccat gtgacggaga
     4021 aattaaaccg gatcaaaaaa gcccagcgca aaattgcgca aacaaagggc agaactgcca
     4081 accttgagga gattgctcaa gaacttgaga tgacccccga tcaagtgcgg gaagtcctgt
     4141 tgcgggtgcc caaatcgatt tccctagaaa cccgtgttgg caaggaaaag gatacggagc
     4201 taggggattt gctggaaacc accgatttca ccccagaaga acgattgact caggaatccc
     4261 tcagcaatga tctgcagcgg ttattgctcg aactcaatca ccgcgaacgc gatgtaattc
     4321 ggatgcggtt tggtctgcgc gatgggcaga tttattccct cgctgaaatt gggcaagtgc
     4381 tcgacctatc gcgtgagcga gttcgccaga ttgaggctaa ggccctgcaa aagctgcggc
     4441 atccgcgacg acgcaatcag atccgtgact atttggagtt gttgcacggc agcggcagcg
     4501 attacaaaga tcacgatggc gattacaaag atcacgatat cgattacaaa gatgatgatg
     4561 ataaatagac ccgggctcga gctagcaagc ttctctagag ctttatgctt gtaaaccgtt
     4621 ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt aattagtaat
     4681 ataatctatt aaaggtcatt caaaaggtca tccaccggat caattcccct gctcgcgcag
     4741 gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc ttggagccct tgccctcccg
     4801 cacgatgatc gtgccgtgat cgaaatccag atccttgacc cgcagttgca aaccctcact
     4861 gatccgcatg cccgttccat acagaagctg ggcgaacaaa cgatgctcgc cttccagaaa
     4921 accgaggatg cgaaccactt catccggggt cagcaccacc ggcaagcgcc gcgacggccg
     4981 aggtcttccg atctcctgaa gccagggcag atccgtgcac agcaccttgc cgtagaagaa
     5041 cagcaaggcc gccaatgcct gacgatgcgt ggagaccgaa accttgcgct cgttcgccag
     5101 ccaggacaga aatgcctcga cttcgctgct gcccaaggtt gccgggtgac gcacaccgtg
     5161 gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg
     5221 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     5281 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     5341 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     5401 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc
     5461 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     5521 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     5581 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     5641 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     5701 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     5761 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     5821 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     5881 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     5941 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     6001 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     6061 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     6121 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     6181 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     6241 atgtctaaca attcgttcaa gccgacgccg cttcgcggcg cggcttaact caagcgttag
     6301 atgcactaag cacataattg ctcacagcca aactatcagg tcaagtctgc ttttattatt
     6361 tttaagcgtg cataataagc cctacacaaa ttgggagata tatcatgaaa ggctggcttt
     6421 ttcttgttat cgcaatagtt ggcgaagtaa tcgcaacatc cgcattaaaa tctagcgagg
     6481 gctttactaa gctgatccgg tggatgacct tttgaatgac ctttaataga ttatattact
     6541 aattaattgg ggaccctaga ggtccccttt tttattttaa aaattttttc acaaaacggt
     6601 ttacaagcat actagaggat cggcggccgc ggatccccgg gtaccgagct cgaatttcga
     6661 gcttctggag caggaagatg tcgcgggcat tagcaccagc ggtctgccaa gcctccgcca
     6721 gccgttgggt cccttccgct tgagcttttc catcttcgac gatacgggcg gcggcccccc
     6781 gcgcttccgc gatcgcccgt ttacaagctg cctcagctgg ggcgatcaca tcggcttgaa
     6841 gttgctgctg cacctgtttg atccgctcct gctgcacagg gagttctgct tggctacgag
     6901 cgacttcggt agcaatgtcc gcttcagctt cggccaccac cgcttcgcgc cgcgtcaacg
     6961 catcctgaat ccggcgctcg gcctcggctt gggcgatcgc tacatcgcga tcgatccgac
     7021 gcagggccgt gatcttgtca ttttcggccg tttggatcgc agaggcagcc tgggcatcgg
     7081 cttcagcaat tcgggcatct cgctgcagat cagcccgctg cttgcgtcca ctagccgaga
     7141 gataaccgac ctcatcggaa atgttctgga cttgcagcgt atcgaggact agacccagct
     7201 gctcaaggtc atcctccgcc tcttccagca gacttttggc aaaggcaatt ttgtcctcgt
     7261 tgatctgctc cggcgtgagg ctggctaaaa caccacgcaa gttgccttcg agggtctcct
     7321 tggcaatttg ctcgatttcc ttacggtttt tgccaagcag ccgctcgatc gcgttgtgga
     7381 tggtcggttc ttccccagca atcttgatat tggcaacgcc ttcaacagtc aggggaatgc
     7441 cgcccttgga gaaggcattg gaaacgcgca actcaatgat catgttggtc agatccatgc
     7501 ggagcgcttt ttccagcaga ggtacccgca ggctgctgcc gcccttgacc aagcgatagc
     7561 caactcggcg gccatcacta ctgcggcgac tactgccagc aaagatcaaa atttcactgg
     7621 gttggcagat gtagtagaga ttgcgcagga ctaagctgcc agccccggcg
//
