LOCUS       sequence-232073-        9881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RpoD6 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 9881)
  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 9881)
  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: KFP06
FEATURES             Location/Qualifiers
     source          1..9881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..109
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(136..141,142..158,159..164)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             3131..3196
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      complement(3665..3692)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3784..3870)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(4244..4249,4250..4266,4267..4272)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4461..4994
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(6084..6103)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             7561..7752
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(7746..7768)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7854..7994
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7909..7928
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8009..8026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8180..8768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8260..8279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8939..9730,9731..9799))
                     /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     9562..9581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9800..9881,1..23))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
       61 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
      121 caagaattct catgtttgac agcttatcat cgataagctt taatgcggta gtttatcaca
      181 gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc tcatcgtcat
      241 cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg tactgccggg
      301 cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg tgctgctagc
      361 gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt ccgaccgctt
      421 tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact acgcgatcat
      481 ggcgaccaca cccgtcctgt ggatcccagg ctcagtgtgg ttcgcatctt ttgatgatca
      541 ccactggacg caacacagta gggcaattca agagcatcca aagcctcttc aaccccagga
      601 acggcttgta gatgcgtttc taacgcgata cgggtacggc gttgatagtg ctgaacaaag
      661 tcagggggtg gaggattgcc taaccgtcgc tcaattagtt tgagacagtc agccatggaa
      721 tgacccacaa actgctcaaa catgtcatcc aaagtcacca acagacccag ttcattgagc
      781 atgtctgcaa agacgcgatt agtgatgcgt tcgctatcaa caagcacacc atcacagtcg
      841 aaaatcacca gctgaaacgg tgaagtttgc attgttttta agcacgagcc atcaacagta
      901 agagcgatcg cgctgggacg atgtaatcgc gccgtaggca gggttttgcc tcatccggca
      961 gatgacaagc ttcaagattc ggcagtgaag tatcgagcaa gcgataccaa acgcggccgc
     1021 gaggaggcct cggcaactgg aagcgcaggt cttcccagta agcattaaag gctaggtaaa
     1081 gccattcctg ctggcgagga tggcagagac tgacggccag actgtgggac cacagcgccc
     1141 aatcgggttg tttgagtttg acgccatgcc agatggcata gggacgacgc ggatggggtt
     1201 cgttctgcag cagttcgttc tgttggaaca tcaccagcga ctgggaaagt tcaatcaggc
     1261 ggcgactgaa caccaagaaa tcggcatggc gatcgcacag cgaccaatca aaccagctga
     1321 tctcattgtc ttggcagtag gcgttattgt taccctgctg actgcgtttg acctcatcgc
     1381 ccatcgtcag catcggtgtg ccctgggcga ggaataacgt ggcgagcaaa ttgcgctgct
     1441 gccgttcccg taagctcaga atcgtggggt catcggtctc tccttcaatg ccgtagttcc
     1501 agctgtagtt gtcattggtc ccgtcccgat tgttctctcc attggcaaag ttgtgcttct
     1561 ggctatagct gactcgtcac gggtaaccga tatcgtcgac aggcctaggt cagaagtgga
     1621 gttcagcatc gccccacttt agcaagacaa aagggcaaga ccctaggccc cgcccaacag
     1681 cgactgagac cagcggtcgc aatcgaatga gtaacgagat tgtaaaaaac tgatgatgcg
     1741 atcgttgatt tcgtgatgtt gagaggtctt actttttaat cactaaaatc aacaatattt
     1801 atcaaggcaa ctatttcgag acactgattt gggtttgacc gcgagtgctc cctagccgat
     1861 cgataacccg ccactgctgc tctttcggct ggtagagctt ttgcaggaac tcaagcgccg
     1921 actggcggtc gcaaaactct aagaattgac cattcacttc tacgaagaaa ggcgcgggta
     1981 acttgtagtc agccatacag aaatttttca gaactcagca gaatgcccca aggatggcag
     2041 aacttcgtta agctgtagtt atttaaaaat catggagaag tgaagaaact gtaatccctc
     2101 ttacattttc gacacattcc tgatctgaag ataaccttaa gagccttggt taaaatcaac
     2161 ttcttgctgc atttttctct taatgctttc gggggtgcgt atggcgagtg cttcggctca
     2221 tgctgtcgcg acggctcagc gacgaacaag agataacgtc agttggtatc tcagctccat
     2281 cggtcggatt ccattgctca ccgccgatca ggagatcgag ctggggcaga aagtgcaggt
     2341 gatgatgaac ctgctggaag aaacttctag cgatcgcacg tttactgagg ccgaaaccgc
     2401 agcgatccag ctggggcagc gtgctaagga acggatgatc aaggccaact tgcgtcttgt
     2461 ggtctctgtg gccaaaaagt accaaggtaa gggtcttgag tttcttgacc tgatccaaga
     2521 aggcgctctc gggctagaac gtgctgtcga gaaatttgac ccctccaagg gctacaagtt
     2581 ttccacctat gccttttggt ggattcgcca gagcatgaca agggcgatcg cctgccaatc
     2641 gcgcactatt cgcctgccgg ttcacttgac tgaacggctg agccaaatcc gcaaagtgac
     2701 tcgggaactg gcgcacgagc taggccgttt gccctctcgc caagaaattg cagaagcggt
     2761 tcagttgcca ctgaatgacc tcgaaacaat tctgcgtcag tcgcgccaat gcaccagcct
     2821 agatgctcca gtcagtggtg aagatggcaa gagtttcctg ggtgagctga ttgctgatcc
     2881 caacggcaat gagcccatgg acatcatgga aaatgccatg cagcgcgagc agcttgatgc
     2941 ttggctcgag catctcacgc cccaagagcg ccaggtgctc tatctccgct ttggactggg
     3001 tgatggcgaa cgtcataccc tcgctgaaat cggccggatg atggaagtct cgcgcgagcg
     3061 ggtccgccaa gttgaggtca aagcgctgcg taagctccgc aacctgacga agcgatcgcg
     3121 gattgctgtc gactacaagg accacgacgg tgactacaag gaccacgaca tcgactacaa
     3181 ggacgacgac gacaagtgag tcgcttccct gtccagtcct cctgtctgtc ttaagcagct
     3241 taggcgcgat cgcctaggct gtttttttga ccgtccattg cgtgatcggt gccagcgatt
     3301 tccagccgag aaagcggccg atcggttccg ctcgctgcac cgccacacgg attaaatctc
     3361 ccccccagcg ctgctgtcca gccaaaacga cttgctcgct ttcgagggta acggcgttaa
     3421 caactagccg cccacccatc ggcagcgctt gccagcattg atcgagcaca cctggagcag
     3481 tgacaccgcc accgatgaag atggcatccg gttggggcaa gccactcagc gtagccggtg
     3541 ctgaaccggc aaccacttgc aggttcggaa ccccgagggc gatcgcattc tgtgtaatga
     3601 ttacccgggc tcgagctagc aagcttggcc ggatccggcc ggatccggag tttgtagaaa
     3661 cgcaaaaagg ccatccgtca ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg
     3721 cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg
     3781 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     3841 gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcatggg
     3901 gagaccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc atggggtcag
     3961 gtgggaccac cgcgctactg ccgccaggca aattctgttt tatcagccgt taccccacct
     4021 actagctaat cccatctggg cacatccgat ggcaagaggc ccgaaggtcc ccctctttgg
     4081 tcttgcgacg ttatgcggta ttagctaccg tttccagtag ttatccccct ccatcaggca
     4141 gtttcccaga cattactcac ccgtccgcca ctcgtcagca aagaagcaag cttagatcca
     4201 ctagtgacgc acaccgtgga aacggatgaa ggcacgaacc cagttgacat aagcctgttc
     4261 ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     4321 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     4381 tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     4441 gatgttatgg agcagcaacg atgttacgca gcagcaacga tgttacgcag cagggcagtc
     4501 gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc
     4561 ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagacg
     4621 tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta
     4681 agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt
     4741 acgttctgcc caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct
     4801 ccggcgagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg
     4861 ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag
     4921 tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa
     4981 gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgcg gagttgttcg
     5041 gtaaattgtc acaacgccgc actagtggat ctggtaaccc cagcgcggag atctcgcagc
     5101 gtaaagccgt catggcaggt gatgaagtta atggtgcgtc cggcatacca ttggtctgtg
     5161 ctgtagacat cggggctacc cagcaggcgt tgactgaggg cgtaagtaca gccctgatct
     5221 ccacgccaaa aacgccgaat atcgtcccgg aagggaccgt tccaagtccc aaagcgatcg
     5281 ccaataaagg taccaacctg atataagccg gctgcgtccc aagcttcagc aatgagcttc
     5341 gtaccggcca aaaccggatc ggaatcaatc gcccaaagca agggcggatc cgataggggg
     5401 ttgccattgg catcacgact cagcaccgac gcaaggtcaa agcggaagcc atcgacgtgc
     5461 atttccgaga cccaataacg caggcaatcg agaatcagcg atcgcacgat cgaattgtta
     5521 gctttgacgg tgttgccgca gccggtgtag ttgcgatagc ccgcctgttc atccagcaga
     5581 tagtaggttg aattcgctag acctttgaaa gacagcgttg gaccgtcttc attcccttca
     5641 gcagtgtggt tgaacaccac gtcgagaatc acctcaatcc ctgcttggtg gagcgccttg
     5701 acgaggtcgc ggaactcatc aactggacca agtggatcat ggcgagagct gtaggctgcg
     5761 tggggcgcaa aaaaagccat ggtgctgtag ccccagtagt tctcgcgtcc taagggggca
     5821 tcttggcgat cgaactggtg caccggcagc aactcaacgg ccgtgacgcc gagggattgc
     5881 aggtagggaa ttttttcgat tagaccagcg taggtgccac gtttttcagg ggcgacgccg
     5941 gaattgggat gcttggtgaa gccgccaaca tgcagctcat agattactgt gcgagcgtag
     6001 ggtgtgcgtg gatggcgatc ggatcctcta cgccggacgc atcgtggccg gcatcaccgg
     6061 cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc
     6121 tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
     6181 cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa
     6241 cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg
     6301 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     6361 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     6421 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     6481 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     6541 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg
     6601 ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct
     6661 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
     6721 cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat
     6781 tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc
     6841 atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg
     6901 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact
     6961 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg
     7021 cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc
     7081 gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc
     7141 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
     7201 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
     7261 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc
     7321 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg
     7381 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc
     7441 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc
     7501 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca
     7561 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
     7621 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
     7681 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
     7741 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     7801 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     7861 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     7921 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     7981 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     8041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     8101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     8161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     8221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     8281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     8341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     8401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8581 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     8641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     9001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     9061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     9121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     9181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     9241 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     9301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     9361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9541 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9841 attgtctcat gagcggatac atatttgaat gtatttagaa a
//
