LOCUS       sequence-201978-        8755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  drives expression of kaiBC from an rpaA-independent promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8755)
  AUTHORS   Puszynska AM, O'Shea EK
  TITLE     Switching of metabolic programs in response to light availability is
            an essential function of the cyanobacterial circadian output 
            pathway.
  JOURNAL   Elife. 2017 Apr 21;6. pii: e23210. doi: 10.7554/eLife.23210.
  PUBMED    28430105
REFERENCE   2  (bases 1 to 8755)
  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"; journalName: "Elife."; 
            date: "2017-04-21"; volume: "6. pii: e23210. doi"; pages: " 
            10.7554/eLife.23210"
COMMENT     Sequence Label: P0050::kaiBC
FEATURES             Location/Qualifiers
     source          1..8755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1110..1925)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1110..1925)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1833..1852
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1917..1936
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4583..5773
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(4935..4954)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             6412..6603
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6597..6619)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6705..6845
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6760..6779
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6860..6877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7031..7619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7111..7130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7790..8581,8582..8650))
                     /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     8413..8432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8651..8755)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctcat gtttgacagc
      121 ttatcatcga tagtgctctc gcaaattgca atctgcattt gggtggaatc gacggcaatt
      181 ttgcaggatt gccagcgggc gctgtcggcc gatcgctatc aactccaagt ctgtgagtct
      241 ggcgaaatgc tcttggagta tgcccaaacc catcgtgacc aaatcgactg cctgatttta
      301 gtggcagcca atcccagctt cagggcagtt gttcagcagc tctgctttga gggagtggtg
      361 gtaccagcga ttgtcgtagg cgatcgcgac agtgaggatc ccgatgaacc agccaaagaa
      421 cagctctatc acagcgctga actgcacctc ggtatccatc agctcgagca attgccctac
      481 caagttgatg ctgcactggc tgaatttctg cgcttagccc cggtcgagac catggccgac
      541 cacatcatgc tgatgggggc caaccacgat cccgagctat cgagccagca gcgggacctc
      601 gctcagcgac tacaagagcg cctaggctat ctcggggtct actacaagcg tgatcccgat
      661 cgctttctgc gcaacctacc cgcctacgaa agccaaaagc tgcaccaagc gatgcagact
      721 agctatcgtg aaatcgtttt gagctatttt tcgccgaata gcaacctcaa ccagagcatt
      781 gacaacttcg tcaacatggc tttctttgcc gatgttccag tcaccaaagt ggtagaaatt
      841 cacatggagc tgatggacga gtttgccaag aagctccgcg tagagggacg ttcagaggac
      901 attttgctgg attatcggct gactttaatt gatgtaattg cacatctttg tgagatgtat
      961 cgacggtcta tcccacgaga aacctgaaaa ggtaaaggag gtcttaagct cggctcaatt
     1021 tctctcttta tcctgttaga tggtttgatt gctgttgcta ccccgttgat ctgcgtctgc
     1081 cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     1141 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1201 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1261 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1321 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     1381 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1441 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1501 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1561 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1621 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1681 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1741 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     1801 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1861 tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg
     1921 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     1981 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     2041 ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcaggg
     2101 ccgcgctgcc tttggttgca ataacggtaa atttaccgtc tgtcccgatt gtgccagtca
     2161 cgttaatatt cgctgcttta ccacttccag cagccgttac ttggctagca ttgccgcaag
     2221 ttatgttaaa gttacctgca ctgctaccaa taccaagctg tgtacaagac gcattggtga
     2281 cgtagttagg atctagtgtc gccgcactaa gggtacgaag agcagcaccg actgctgatt
     2341 tgccttcaga ggcataggct ttatctgttt gatccaacag gttcggtaat gcaactgctg
     2401 ccaagatgcc aacgataatg ataacgacca gcaactcgac aagggtaaaa cctttttctt
     2461 cttcaccttg ttcgtcttta cgctgggcaa gatattgtag ataacgaaga cggagggagt
     2521 cagacattca ggagatcctc agcaagagag ctgttcttga agtaacttac ccctacagca
     2581 ggcttttatt tcaggttcct gaaaatttcc gcatgagccc tcgtaaaacc tacattctca
     2641 agctctacgt cgccggcaat actccaaact cagtccgtgc cctcaaaacg ctcaagaaca
     2701 ttctcgaagt tgaatttcaa ggtgtttatg ctctaaaggt gatcgatgtt ctcaaaaatc
     2761 ctcagttggc agaagaggat aaaatcctag cgacgccaac cctcgccaag gttctaccac
     2821 tgcctgtccg acggattatt ggtgatttat ccgaccgtga gaaagttttg attggccttg
     2881 atttactcta cggcgaactt caagattccg acgacttcta actgtcgtta actgctttgt
     2941 tggtactacc tgacttcacc ctcttttaag atgacttccg ctgagatgac tagccctaat
     3001 aataattctg agcaccaagc catcgctaag atgcgcacga tgattgaagg ctttgatgat
     3061 attagtcatg gcggtcttcc aatcgggcga tcgaccctcg ttagtggtac ttcaggaacc
     3121 ggcaagaccc ttttttctat tcaatttctc tataacggta ttatcgagtt tgatgagcct
     3181 ggggttttcg ttactttcga agaaaccccg caagatatca ttaaaaacgc ccgtagtttt
     3241 ggctgggatt tagccaagct ggtcgatgag ggcaaactat ttattcttga tgcttcaccc
     3301 gatccagaag gtcaagaggt tgttggcggc ttcgatctct ctgctctgat tgagcggatt
     3361 aattatgcaa ttcaaaagta tcgagcgcgg cgggtttcaa ttgactcggt cacgtccgtt
     3421 ttccagcaat atgatgcctc ttctgtggtt cgccgcgaac tctttcggtt ggtagctcgc
     3481 ctaaaacaaa ttggggcaac tacggtcatg accaccgagc gtatcgagga atatggcccg
     3541 atcgctcgtt acggtgttga ggaatttgtc tccgataacg tcgtgattct ccgcaacgtt
     3601 ttggaagggg agcgccgtcg ccgcaccctc gaaatcctca agctacgtgg caccagccac
     3661 atgaaagggg aatatccgtt cacgattacg gatcatggca tcaatatctt cccgctcggg
     3721 gcaatgcgcc ttacgcagcg atcgtcgaac gtgcgtgttt catctggtgt cgtccgactc
     3781 gatgaaatgt gtggtggggg cttctttaag gactcaatca ttctggcaac tggcgctaca
     3841 ggcactggta aaactctgtt agttagccgt ttcgttgaga atgcttgtgc taacaaagag
     3901 cgggcgattc tgttcgctta tgaagagtca cgagctcagc tgctccgcaa tgcctattca
     3961 tggggaatgg actttgagga gatggagcgc caaaacctcc tcaaaattgt ttgcgcctat
     4021 cctgaatctg caggtcttga agaccatttg cagattatta agtcggagat caatgacttt
     4081 aagccagctc gtattgcaat cgactccctc tctgctttgg cgcggggcgt tagcaacaat
     4141 gccttccgcc aatttgtaat tggtgtcact ggctacgcga aacaagaaga aatcacggga
     4201 ctattcacaa ataccagtga tcaatttatg ggagcgcatt cgattactga ctcccatatc
     4261 tcaacaatta cggatacgat tatcttgctc caatacgtcg agattcgtgg cgaaatgtcc
     4321 cgcgccatta acgtcttcaa gatgcgcgga tcttggcatg acaaagcaat ccgcgaattc
     4381 atgatcagcg acaaagggcc ggacatcaag gattctttcc ggaactttga gcggattatt
     4441 tcaggttcgc caacacggat taccgtcgat gagaaaagcg aactctcgcg aattgtgcgc
     4501 ggcgttcaag aaaaagggcc ggagagctag ttaatgcggt agtttatcac agttaaattg
     4561 ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca tcctcggcac
     4621 cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg
     4681 ggatatcgtc cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc
     4741 gttgatgcaa tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg
     4801 cccagtcctg ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac
     4861 acccgtcctg tggatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     4921 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     4981 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     5041 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     5101 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gaccgatgcc
     5161 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     5221 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg
     5281 ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc
     5341 ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg
     5401 tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt
     5461 gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg
     5521 cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca
     5581 gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca ttggaccgct
     5641 gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt
     5701 aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc
     5761 cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt
     5821 ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata
     5881 tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc
     5941 tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
     6001 tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc
     6061 tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa
     6121 agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga
     6181 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc
     6241 tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt
     6301 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc
     6361 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa
     6421 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg
     6481 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac
     6541 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6601 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6661 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     6721 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     6781 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6841 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6901 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6961 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7021 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7081 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7141 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7201 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7261 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7321 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7381 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7441 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7501 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7561 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7621 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7681 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7741 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7801 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7861 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7921 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7981 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8041 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8101 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8161 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8221 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8281 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8341 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8401 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8461 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8521 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8581 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8641 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8701 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcg
//
