LOCUS       sequence-232104-        5251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rpoD2∆::NatR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5251)
  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 5251)
  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: KFP79
FEATURES             Location/Qualifiers
     source          1..5251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2851..2868)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2851..2867)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2860..2882)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3076..3095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3195..3217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3255..3273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3341..3445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3446..3514,3515..4306)
                     /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     complement(3664..3683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4477..5065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4966..4985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5219..5236
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattc gagctcggta
      241 cctgcactgg tactacatct tccagaccaa tcaagccgcg ctagaagctg cccaaaaaca
      301 cgacatgggt gtgttcatca tcagcccctc tgataagggc ggacacctct actcaccgcc
      361 gcaaaaactg gtcgatctct gcgagccact gcaccccatg gtcttcaatg acctcttttg
      421 cctcagccag ccccaaatcc ataccctcag cattggtgcg gcgcggccca gtgattttga
      481 tcgccacttg gaaaccttgc ccttgcttga ccaagcggat caggtattgc cgccaatctt
      541 ggagcgcctc gaagcagctg ccactgctgc actggggcaa gactggctgg acacctggat
      601 ggttggcttg cccacccctg acaccacccc gggggaaatt aatatcccga cgattgtctg
      661 gctttacaat cttgtgcaag ctttcgacat ggtcaaatac gccaaagctc gctacaactt
      721 gcttggtaat ggtggccact ggtttccggg gcaaagggca ggacagtttg atccagttgc
      781 actctccgca gccctcgatc aaagccctca tcgcgatcgc attctccaaa ttctcaagga
      841 aatggatgct tggttggggg gtgagtctgt cgttcgactc tcccaaagca gctgatcctt
      901 aagacgatcc cctgcgcgat cgcccagact ttagggaact ttaaggatac ttacaacgtc
      961 ctgctaggca gtgaagcttt taagggtgcc tagtcatgtc agcttgcgcc actacagcca
     1021 agaagctcat tccggttttc cctacctagt cagtcaaact cataacgatt atgatttggc
     1081 catattgctc tcccttaaaa aaaagcccgc tcattaggcg ggctggccgg ccttaggggc
     1141 agggcatgct catgtacagg gcctgttcgc catcactggc ggtgccatcg tacagagcgg
     1201 tatccaggcc acacagggtg aagcccatgc ggcgataggc gtggatagcg ggagcgttca
     1261 cgttggtcac ttccagccac aggtggccag caccgcgttc gcgagcaaat tcggtggcca
     1321 ggcccatcag agcgcgacca acgccgtgac cgcggtgttc gggagccact tcaatatctt
     1381 ccacggtcaa gcggcgattc cagccgctgt agctcaccac cacaaagcca gccagatcgc
     1441 catcatcacc gtaggccaca aaggtgcggc tatcgggatc gccatcttcg ccagcatccg
     1501 attcatcatc actttcatca tcgggaaaca ctttggtcag agggggatcg acgggcactt
     1561 cgcgcagggt aaagccatca cccgtggcgg tcacgcgaaa cacggtatcg gtggtaaagc
     1621 tgccatccag ggcttcgatg gcttcggcat caccgggaac gctggtgcgg tagcggtagg
     1681 cggtatcatc cagggtggtc atatggatcc ggccaccgtt tgcttttaac gtcagccttt
     1741 tacctaaatt cccatcaagc gatcttttaa tatcggacaa cgtcatcgcc attgcatcca
     1801 cctcactaca tttattgtac aacacgagcc catttttgtc aaataaaatt taaattatat
     1861 caacgttaat aagacgttgt caataaaatt attttgacaa aatttccttt gcttgtttgc
     1921 tctattccta taattcgatg ataagctgtc aaacatgaga attcaacttg catgttcagg
     1981 ttttgtcaca accgcaaact gggatggttt aatgggctca agcttgcaaa aagtagcaaa
     2041 taatacaaaa tggcaccgcg atcgctcaga cggctgcatt ggcattggga gcaacaacga
     2101 tcgcaatcag acctgattca gccccgctac tctcggcaac aagtgctggc atcttggctc
     2161 ggagccctcg tcggtattgg tcttttgggc tggttgaccc aagcgagtgg ctaccctttg
     2221 gtagcagctc ccatgggtgc gacctcggtg ttgctgttcg gcttacccaa tagtcctttg
     2281 gcgcagccgc gcaatgtcat tttgggcaat gggctcggtg ctctaattgc tgttctgtgt
     2341 gtgttgctct tgggtgctaa cccttggagc atgggctttg ctgtcggtgt cacgatcgct
     2401 ctcacccaat tgttgcgctg tgtccatcct ccggcgggtg cagttgctct cctcggagtg
     2461 attgtcaaag cgaagtgggc ttacatcctg ctgcctgttt tgagcggatc gattctcctg
     2521 tgcgtgatta cggcgctcta tagtcgctgg gcacccgatc gccatcatcg ccgctatcct
     2581 gtgcactggc tctagctaat cgacgttctt taatcggcag tgatctgcaa ccagtcctga
     2641 agtcgttcgg gcaagggcga tcgctgtctt gtttggcttt ctaggcaaag gtgaatggtt
     2701 tgggctgtgg ccactggctg atggttgtgg ctgagacgat aatcaacccg aaagcgatcg
     2761 cgacccaaag cggtagcctg taactcaatc tctaggcgat cgcgtacccg gggatcctct
     2821 agagtcgacc tgcaggcatg caagcttggc actggccgtc gttttacaac gtcgtgactg
     2881 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2941 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3001 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3061 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3121 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3181 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3241 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3841 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3961 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4021 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4081 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4141 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4201 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4261 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4321 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4381 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4441 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4501 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4561 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4621 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4681 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4741 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4801 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4861 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4921 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4981 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5041 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5101 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5161 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5221 cgagtcagtg agcgaggaag cggaagagcg c
//
