LOCUS       sequence-232097-       10043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RpoD5 epitope tagged construct for integration at neutral site 2.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10043)
  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 10043)
  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: KFP05
FEATURES             Location/Qualifiers
     source          1..10043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1938..2003
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      complement(2069..2096)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2188..2274)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(2648..2653,2654..2670,2671..2676)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2865..3398
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(4488..4507)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5965..6156
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6150..6172)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6258..6398
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6313..6332
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6413..6430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6584..7172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6664..6683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7343..8134,8135..8203))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7966..7985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8204..8308)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8376..8394
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8421..8426,8427..8443,8444..8449)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 acagagtttt ggtaatgaca gcttctgtag actcgcgctt ctgtccatca tgccaagact
       61 ctaaacctgc tcgagaagca ccagcgaaaa ccagttttgc tcacagattt tacgagctgc
      121 tgcgtcatac agggaaatga agagcatgaa gcgctaaaga gttccgaccc attcctggga
      181 attgcgggga acctccatca tactcggcgc tgtcgagaaa caagaataca gccaagcggg
      241 gctggacgcc gcattgagaa agatttttgc agctctaaca gccccaagcc agcaggcaag
      301 caccctttcc agcgccccca gagcccagag gctcactcat agaatcacga ttctcctagg
      361 gcaaagccgc aatggccctc taggctggat tccgagtagg ggagtcattt cagacgagcg
      421 atcgccaaca atgcaaactc agcgttgatg ccacatgttt tctcagaatt cttgatggtt
      481 ctcaatggtg agctagaggg atacaaaagt tcaagaaacc aagcactgaa ctgggcaaat
      541 atcaaagcaa catctgggct ttaccgagga cttacatttt ggccaaaatc taaagagagc
      601 tggggcactg gcaagaattt tattttttat agagagacta ttttccagat aagcactggc
      661 cttgaagtcg acggcaaaat attgcatcgc tccagccagt cgttatccct gtaataaact
      721 ggatattcac catgacggct gcctatttca ctcattccac ggaatacgaa gcagtttcgt
      781 ctgacgtcgc taacttgtca gaaactccaa gttctgaagc cgtggaaaca gcggtctcta
      841 gccgccagtc aaccgatctg attcggctgt atttacaaga catcgggcga gtccgtctcc
      901 ttgggcgcga tgaagaggtg agtgaggcac ggtgggtgca gcgctacatg cagttgttgg
      961 cggttcggaa ttgcgtcgcc gttgaagaag gaggggttct caaggattac gttgacctgc
     1021 aggaaggccg cgatcgcctg accgcgcagc tgggtcaccg tccttccttt gaagcttggg
     1081 ccaagattgt cgattgttcc ttgactgaat tacgccaagc actacaactg gggctgcatc
     1141 gctgggctga agtctgccag ctcgatcttg aggaattaaa gcaggtctgc aaagaaggcc
     1201 agcgagctaa agatcacatg atcaaggcca acctccgttt agttgtttcc gtcgctaaaa
     1261 aatatcaaaa ccgaggacta gagcttctcg atctcgttca agaaggcacc ctaggactag
     1321 agcgggctgt tgagaagttc gacccgacga aaggctatcg atttagtacc tatgcctatt
     1381 ggtggattcg tcagggaatt acccgcgcga tcgcaaccca aagtcggacg attcgcttgc
     1441 ccgtccatgt gacggagaaa ttaaaccgga tcaaaaaagc ccagcgcaaa attgcgcaaa
     1501 caaagggcag aactgccaac cttgaggaga ttgctcaaga acttgagatg acccccgatc
     1561 aagtgcggga agtcctgttg cgggtgccca aatcgatttc cctagaaacc cgtgttggca
     1621 aggaaaagga tacggagcta ggggatttgc tggaaaccac cgatttcacc ccagaagaac
     1681 gattgactca ggaatccctc agcaatgatc tgcagcggtt attgctcgaa ctcaatcacc
     1741 gcgaacgcga tgtaattcgg atgcggtttg gtctgcgcga tgggcagatt tattccctcg
     1801 ctgaaattgg gcaagtgctc gacctatcgc gtgagcgagt tcgccagatt gaggctaagg
     1861 ccctgcaaaa gctgcggcat ccgcgacgac gcaatcagat ccgtgactat ttggagttgt
     1921 tgcacggcag cggcagcgat tacaaagatc acgatggcga ttacaaagat cacgatatcg
     1981 attacaaaga tgatgatgat aaatagaccc gggctcgagc tagcaagctt ggccggatcc
     2041 ggccggatcc ggagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     2101 aatttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     2161 cgcaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     2221 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     2281 ggcagttccc tactctcgca tggggagacc ccacactacc atcggcgcta cggcgtttca
     2341 cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaattct
     2401 gttttatcag ccgttacccc acctactagc taatcccatc tgggcacatc cgatggcaag
     2461 aggcccgaag gtccccctct ttggtcttgc gacgttatgc ggtattagct accgtttcca
     2521 gtagttatcc ccctccatca ggcagtttcc cagacattac tcacccgtcc gccactcgtc
     2581 agcaaagaag caagcttaga tccactagtg acgcacaccg tggaaacgga tgaaggcacg
     2641 aacccagttg acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc
     2701 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     2761 catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc
     2821 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca
     2881 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca
     2941 tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc
     3001 ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt
     3061 acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag
     3121 cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga
     3181 tctatatcta tgatctcgca gtctccggcg agcaccggag gcagggcatt gccaccgcgc
     3241 tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag
     3301 cagattacgg tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag
     3361 tgatgcactt tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg
     3421 gcttcccggc cgcggagttg ttcggtaaat tgtcacaacg ccgcactagt ggatctggta
     3481 accccagcgc ggagatctcg cagcgtaaag ccgtcatggc aggtgatgaa gttaatggtg
     3541 cgtccggcat accattggtc tgtgctgtag acatcggggc tacccagcag gcgttgactg
     3601 agggcgtaag tacagccctg atctccacgc caaaaacgcc gaatatcgtc ccggaaggga
     3661 ccgttccaag tcccaaagcg atcgccaata aaggtaccaa cctgatataa gccggctgcg
     3721 tcccaagctt cagcaatgag cttcgtaccg gccaaaaccg gatcggaatc aatcgcccaa
     3781 agcaagggcg gatccgatag ggggttgcca ttggcatcac gactcagcac cgacgcaagg
     3841 tcaaagcgga agccatcgac gtgcatttcc gagacccaat aacgcaggca atcgagaatc
     3901 agcgatcgca cgatcgaatt gttagctttg acggtgttgc cgcagccggt gtagttgcga
     3961 tagcccgcct gttcatccag cagatagtag gttgaattcg ctagaccttt gaaagacagc
     4021 gttggaccgt cttcattccc ttcagcagtg tggttgaaca ccacgtcgag aatcacctca
     4081 atccctgctt ggtggagcgc cttgacgagg tcgcggaact catcaactgg accaagtgga
     4141 tcatggcgag agctgtaggc tgcgtggggc gcaaaaaaag ccatggtgct gtagccccag
     4201 tagttctcgc gtcctaaggg ggcatcttgg cgatcgaact ggtgcaccgg cagcaactca
     4261 acggccgtga cgccgaggga ttgcaggtag ggaatttttt cgattagacc agcgtaggtg
     4321 ccacgttttt caggggcgac gccggaattg ggatgcttgg tgaagccgcc aacatgcagc
     4381 tcatagatta ctgtgcgagc gtagggtgtg cgtggatggc gatcggatcc tctacgccgg
     4441 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     4501 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     4561 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
     4621 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     4681 ggagtcgcat aagggagagc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     4741 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     4801 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
     4861 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
     4921 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
     4981 cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat
     5041 ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc
     5101 catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc
     5161 tcttaccagc ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc
     5221 ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct
     5281 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
     5341 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
     5401 gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc
     5461 cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc
     5521 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     5581 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     5641 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     5701 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     5761 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     5821 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     5881 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     5941 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     6001 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     6061 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     6121 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6181 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6241 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     6301 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     6361 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     6421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7021 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7261 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7321 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7381 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7441 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7501 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7561 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7621 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     7681 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7741 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7801 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7861 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7921 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     7981 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8041 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8101 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8161 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8221 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8281 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8341 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8401 gtcttcaaga attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta
     8461 tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc
     8521 gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg
     8581 ccgggcctct tgcgggatat cgtccattcc gacagcatcg ccagtcacta tggcgtgctg
     8641 ctagcgctat atgcgttgat gcaatttcta tgcgcacccg ttctcggagc actgtccgac
     8701 cgctttggcc gccgcccagt cctgctcgct tcgctacttg gagccactat cgactacgcg
     8761 atcatggcga ccacacccgt cctgtggatc ccaggctcag tgtggttcgc atcttttgat
     8821 gatcaccact ggacgcaaca cagtagggca attcaagagc atccaaagcc tcttcaaccc
     8881 caggaacggc ttgtagatgc gtttctaacg cgatacgggt acggcgttga tagtgctgaa
     8941 caaagtcagg gggtggagga ttgcctaacc gtcgctcaat tagtttgaga cagtcagcca
     9001 tggaatgacc cacaaactgc tcaaacatgt catccaaagt caccaacaga cccagttcat
     9061 tgagcatgtc tgcaaagacg cgattagtga tgcgttcgct atcaacaagc acaccatcac
     9121 agtcgaaaat caccagctga aacggtgaag tttgcattgt ttttaagcac gagccatcaa
     9181 cagtaagagc gatcgcgctg ggacgatgta atcgcgccgt aggcagggtt ttgcctcatc
     9241 cggcagatga caagcttcaa gattcggcag tgaagtatcg agcaagcgat accaaacgcg
     9301 gccgcgagga ggcctcggca actggaagcg caggtcttcc cagtaagcat taaaggctag
     9361 gtaaagccat tcctgctggc gaggatggca gagactgacg gccagactgt gggaccacag
     9421 cgcccaatcg ggttgtttga gtttgacgcc atgccagatg gcatagggac gacgcggatg
     9481 gggttcgttc tgcagcagtt cgttctgttg gaacatcacc agcgactggg aaagttcaat
     9541 caggcggcga ctgaacacca agaaatcggc atggcgatcg cacagcgacc aatcaaacca
     9601 gctgatctca ttgtcttggc agtaggcgtt attgttaccc tgctgactgc gtttgacctc
     9661 atcgcccatc gtcagcatcg gtgtgccctg ggcgaggaat aacgtggcga gcaaattgcg
     9721 ctgctgccgt tcccgtaagc tcagaatcgt ggggtcatcg gtctctcctt caatgccgta
     9781 gttccagctg tagttgtcat tggtcccgtc ccgattgttc tctccattgg caaagttgtg
     9841 cttctggcta tagctgactc gtcacgggta accgatatcg tcgacaggcc tctaggatct
     9901 gcagtagctc agtgccaact gcaccggtcg ctccaagaat tgcaacgcga tagccctgtg
     9961 acaatgacaa cctcctaacc ccaaacagca tcgatagcga aagcagacaa ctctgcgccc
    10021 attgtctggc ggcgcatcgg cac
//
