LOCUS       sequence-232101-        5191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rpoD6∆::NatR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  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 5191)
  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: KFP75
FEATURES             Location/Qualifiers
     source          1..5191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..113
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     307..329
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     321..338
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     322..338
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2913..2929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2913..2929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2926..2948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2937..2953
                     /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)."
     promoter        complement(join(2961..2967,2968..2985,2986..2991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3006..3027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3144..3161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3315..3903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3395..3414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4074..4865,4866..4934))
                     /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"
     primer_bind     4697..4716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4935..5039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5107..5125
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5163..5185)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
       61 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
      121 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
      181 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
      241 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      301 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtacgattt gtaccgcagc aatctcgaga agatggagca
      421 ggcgcggatc acgatggaag aagccatcca agccgatcgc cgcaagcggg aacagcgcga
      481 gcaggaaaaa ttggcgaaag ccgctgccgc tgaggccccc gcagctctcg aagcctccag
      541 cgacaatccc gagcctgaaa ctagcgaaac gccctcctaa ggcaccaacc gagaccagcg
      601 atcgcatctg actagcgatg tctaaagcga tcctgtgcaa agcaagttcg gctaattccc
      661 cagtgttagc atcgcaggca attgcggccc gaggtttcag gatgcagacc cgaagctgga
      721 cgagtgcagc gatcgccctt ggattgtgct tgggtgtggg ctggagcgca ccagtcttca
      781 gcctgacgcc ttcgcaacgt cagcaactta aaagtcttgg gattccggtg gtgattccca
      841 aggccatccc gacgggtttt caagtcaaaa acgtttccgt caaaccctgc cctgcccaat
      901 ccagtcgtaa cagcaggggc gtctgtcggt ttggcccaga ctacgaaatt ctttaccgca
      961 atagccaagg ggcttgcttc acgatggtgg cggttggggg tgggatcggt ggcgttgacc
     1021 aaacctacgg ctatgaggtc gcagtgccat tgttccgtga gcgcgtcatg ctttggtttg
     1081 gcgacctcaa caccaacact ccttaccgca cgccgacaga gcagcaacga caacaatccc
     1141 agtccaatct acgttccgat tggctgggca aggggccttt ctacggcgtc agctacgtcc
     1201 agagagagcc ccaatgtcgt cggggtatct ccccaaagca ggctgagcag gttttaattc
     1261 tcatgtttga cagcttatca tcgaattata ggaatagagc aaacaagcaa aggaaatttt
     1321 gtcaaaataa ttttattgac aacgtcttat taacgttgat ataatttaaa ttttatttga
     1381 caaaaatggg ctcgtgttgt acaataaatg tagtgaggtg gatgcaatgg cgatgacgtt
     1441 gtccgatatt aaaagatcgc ttgatgggaa tttaggtaaa aggctgacgt taaaagcaaa
     1501 cggtggccgg atccatatga ccaccctgga tgataccgcc taccgctacc gcaccagcgt
     1561 tcccggtgat gccgaagcca tcgaagccct ggatggcagc tttaccaccg ataccgtgtt
     1621 tcgcgtgacc gccacgggtg atggctttac cctgcgcgaa gtgcccgtcg atccccctct
     1681 gaccaaagtg tttcccgatg atgaaagtga tgatgaatcg gatgctggcg aagatggcga
     1741 tcccgatagc cgcacctttg tggcctacgg tgatgatggc gatctggctg gctttgtggt
     1801 ggtgagctac agcggctgga atcgccgctt gaccgtggaa gatattgaag tggctcccga
     1861 acaccgcggt cacggcgttg gtcgcgctct gatgggcctg gccaccgaat ttgctcgcga
     1921 acgcggtgct ggccacctgt ggctggaagt gaccaacgtg aacgctcccg ctatccacgc
     1981 ctatcgccgc atgggcttca ccctgtgtgg cctggatacc gctctgtacg atggcaccgc
     2041 cagtgatggc gaacaggccc tgtacatgag catgccctgc ccctaaggcc ggccagcccg
     2101 cctaatgagc gggctttttt ttccaaggct cttaaggtta tcttcagatc aggaatgtgt
     2161 cgaaaatgta agagggatta cagtttcttc acttctccat gatttttaaa taactacagc
     2221 ttaacgaagt tctgccatcc ttggggcatt ctgctgagtt ctgaaaaatt tctgtatggc
     2281 tgactacaag ttacccgcgc ctttcttcgt agaagtgaat ggtcaattct tagagttttg
     2341 cgaccgccag tcggcgcttg agttcctgca aaagctctac cagccgaaag agcagcagtg
     2401 gcgggttatc gatcggctag ggagcactcg cggtcaaacc caaatcagtg tctcgaaata
     2461 gttgccttga taaatattgt tgattttagt gattaaaaag taagacctct caacatcacg
     2521 aaatcaacga tcgcatcatc agttttttac aatctcgtta ctcattcgat tgcgaccgct
     2581 ggtctcagtc gctgttgggc ggggcctagg gtcttgccct tttgtcttgc taaagtgggg
     2641 cgatgctgaa ctccacttct gacctctcac agccctgtgc tctaaccgat cctgctacta
     2701 atccaacctt gcaagggcgg cctgactcgc ggcaagttga gaaaatccct ggctacacac
     2761 agcccgtcca tgtgatcgct ctatgcgact ggagaactaa gcgatcgcac gtctgcaggt
     2821 tgtgtggggt attcggtcaa gcagctttgg ctgggaattg ggctctcgga aatccgtcat
     2881 gagtgtggta ccgagctcga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     2941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3721 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5041 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5101 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5161 aacctctgac acatgcagct cccggagacg g
//
