LOCUS       sequence-196980-        6530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletes kaiBC locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6530)
  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 6530)
  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: kaiBC-
FEATURES             Location/Qualifiers
     source          1..6530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(1410..1514)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1717..1819
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1820..2479
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1886..1905)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3931..3948)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3931..3947)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3940..3962)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4156..4175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4275..4297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4421..4525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4526..4594,4595..5386)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5557..6145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6046..6065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6299..6316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6433..6454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6469..6474,6475..6492,6493..6499)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6507..6523
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     join(6512..6530,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgacatg attacgaatt cgagctcgag ccgcctttta aacaacagcc
       61 ctctatcatc tcagaaactt ttgagaactg tcaaactgtg gacaaagcga tcgcggcctc
      121 atgagctgca gtgctaggct aaattaaatt tttcctttgt ccagagatta atctgtctgc
      181 agactcagtc ctgacaggag cgactgcgtg ctctcgcaaa ttgcaatctg catttgggtg
      241 gaatcgacgg caattttgca ggattgccag cgggcgctgt cggccgatcg ctatcaactc
      301 caagtctgtg agtctggcga aatgctcttg gagtatgccc aaacccatcg tgaccaaatc
      361 gactgcctga ttttagtggc agccaatccc agcttcaggg cagttgttca gcagctctgc
      421 tttgagggag tggtggtacc agcgattgtc gtaggcgatc gcgacagtga ggatcccgat
      481 gaaccagcca aagaacagct ctatcacagc gctgaactgc acctcggtat ccatcagctc
      541 gagcaattgc cctaccaagt tgatgctgca ctggctgaat ttctgcgctt agccccggtc
      601 gagaccatgg ccgaccacat catgctgatg ggggccaacc acgatcccga gctatcgagc
      661 cagcagcggg acctcgctca gcgactacaa gagcgcctag gctatctcgg ggtctactac
      721 aagcgtgatc ccgatcgctt tctgcgcaac ctacccgcct acgaaagcca aaagctgcac
      781 caagcgatgc agactagcta tcgtgaaatc gttttgagct atttttcgcc gaatagcaac
      841 ctcaaccaga gcattgacaa cttcgtcaac atggctttct ttgccgatgt tccagtcacc
      901 aaagtggtag aaattcacat ggagctgatg gacgagtttg ccaagaagct ccgcgtagag
      961 ggacgttcag aggacatttt gctggattat cggctgactt taattgatgt aattgcacat
     1021 ctttgtgaga tgtatcgacg gtctatccca cgagaaacct gaaaaggtaa aggaggtctt
     1081 aagctcggct caatttctct ctttatcctg ttagatggtt tgattgctgt tgctaccccg
     1141 ttgatctgcg tcccgggatc cgtcacacgg gataataccg cgccacatag cagaacttta
     1201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     1261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     1321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     1381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     1441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     1501 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     1561 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcgaataaat
     1621 acctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg
     1681 gaagccctgg gccaactttt ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa
     1741 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     1801 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     1861 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     1921 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     1981 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     2041 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     2101 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     2161 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     2221 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     2281 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     2341 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     2401 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     2461 gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggttg
     2521 ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac
     2581 ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc
     2641 ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt
     2701 gctcaggctc tccccgtgga ggtaataatt gacgatatga tcgaccccgg gggtttgagc
     2761 ggattatttc aggttcgcca acacggatta ccgtcgatga gaaaagcgaa ctctcgcgaa
     2821 ttgtgcgcgg cgttcaagaa aaagggccgg agagctagcc agcgatcgcg cttaggtatt
     2881 cttgtctcag ctcagtcgtc aacccgagca ccgagaaagc taaggcctcg cgataaagcc
     2941 gttgtgctgg attctgtagc gaattcgccg caccagcggc actgaaaaca gcggcttggg
     3001 tacaacgatg catcagggtc agggcttgaa tgcgtagact cagcgcctct ctgaaatcag
     3061 cttggcccaa ggcttgagcg atcgcctgtt cgagttgccc aagttgggca gttagaactt
     3121 ctcggcaatc tgcatcgcgc gcagtcagca gccctagaca ggcacgtgct aacccacagg
     3181 cgaaggccga gggactgagc gtcccttggc gatcgcgatc gcggatccaa gtggcgggca
     3241 ggatggtcac caagcgatcg cggggcaggg tgatgattag ttgcgctgca accgttccgg
     3301 ttgtacccat cgcggccagt gcgagcggtg agaccttcca ctcaggcgct tgggccggga
     3361 tcaacgcaaa cagtgtttgc ccctctggca atggcgcagc aatcagaaac tcttcaaaaa
     3421 aaccccaacc ggtcagccaa ggcagttctc ccgacagaac gatgtgatcc ttatgccaac
     3481 tggcttgcag gggtgagggc gatcgacgca gatgggaaaa gccaatcccc accgtaggac
     3541 ggccgtagct gaggccagag agtcggcgat cgctgggttg atgcttgctc agaaaagagg
     3601 ctgcactctg gtgctgggtt tgtaaaaaag ccaaggcacc ggaagcactc gcgatcgccg
     3661 cttgagcctg ccatacctca accggtgaga gcgctaaacc ttcctcacag gcagggacag
     3721 aaaacccgag ccaaccgcga tcgcccattg cctgcaacac ctgagccagc actaccggat
     3781 cctgatcgat ccgctcggct tggggctgca cctcagcggt cagccaagct tgcaaatctt
     3841 gcaggtcgaa tcgaggagcc atatcagcgg gcgatcgcga ttaaacctta ggcagcgtct
     3901 agagtcgacc tgcaggcatg caagcttggc actggccgtc gttttacaac gtcgtgactg
     3961 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4021 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4081 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4141 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4201 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4261 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4321 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4381 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4441 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4501 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4561 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4621 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4681 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4741 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4801 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4861 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4921 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4981 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5041 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5101 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5161 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5221 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5281 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5341 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5401 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5461 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5521 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5581 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5641 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5701 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5761 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5821 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5881 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5941 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6001 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6061 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6121 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6181 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6241 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6301 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6361 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6421 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6481 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
//
