LOCUS       sequence-232061-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Psynpcc7942_0456::rpoD2::NatR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  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 6073)
  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: KFP80
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..48)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     148..167
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     361..383
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     375..392
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     376..392
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3849..3865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3849..3865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3862..3884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3873..3889
                     /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(3897..3903,3904..3921,3922..3927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3942..3963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4080..4097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4251..4839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4331..4350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5010..5801,5802..5870))
                     /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     5633..5652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5871..5975)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6043..6061
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
       61 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
      121 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
      181 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
      241 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
      301 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
      361 cccagtcacg acgttgtaaa acgacggcca gtgccaagct tgcatgcctg caggtcgact
      421 ctagaggatc cccgggtaca atcaagccgc gctagaagct gcccaaaaac acgacatggg
      481 tgtgttcatc atcagcccct ctgataaggg cggacacctc tactcaccgc cgcaaaaact
      541 ggtcgatctc tgcgagccac tgcaccccat ggtcttcaat gacctctttt gcctcagcca
      601 gccccaaatc cataccctca gcattggtgc ggcgcggccc agtgattttg atcgccactt
      661 ggaaaccttg cccttgcttg accaagcgga tcaggtattg ccgccaatct tggagcgcct
      721 cgaagcagct gccactgctg cactggggca agactggctg gacacctgga tggttggctt
      781 gcccacccct gacaccaccc cgggggaaat taatatcccg acgattgtct ggctttacaa
      841 tcttgtgcaa gctttcgaca tggtcaaata cgccaaagct cgctacaact tgcttggtaa
      901 tggtggccac tggtttccgg ggcaaagggc aggacagttt gatccagttg cactctccgc
      961 agccctcgat caaagccctc atcgcgatcg cattctccaa attctcaagg aaatggatgc
     1021 ttggttgggg ggtgagtctg tcgttcgact ctcccaaagc agctgatcct taagaattct
     1081 catgtttgac agcttatcat cgaattatag gaatagagca aacaagcaaa ggaaattttg
     1141 tcaaaataat tttattgaca acgtcttatt aacgttgata taatttaaat tttatttgac
     1201 aaaaatgggc tcgtgttgta caataaatgt agtgaggtgg atgcaatggc gatgacgttg
     1261 tccgatatta aaagatcgct tgatgggaat ttaggtaaaa ggctgacgtt aaaagcaaac
     1321 ggtggccgga tccatatgac caccctggat gataccgcct accgctaccg caccagcgtt
     1381 cccggtgatg ccgaagccat cgaagccctg gatggcagct ttaccaccga taccgtgttt
     1441 cgcgtgaccg ccacgggtga tggctttacc ctgcgcgaag tgcccgtcga tccccctctg
     1501 accaaagtgt ttcccgatga tgaaagtgat gatgaatcgg atgctggcga agatggcgat
     1561 cccgatagcc gcacctttgt ggcctacggt gatgatggcg atctggctgg ctttgtggtg
     1621 gtgagctaca gcggctggaa tcgccgcttg accgtggaag atattgaagt ggctcccgaa
     1681 caccgcggtc acggcgttgg tcgcgctctg atgggcctgg ccaccgaatt tgctcgcgaa
     1741 cgcggtgctg gccacctgtg gctggaagtg accaacgtga acgctcccgc tatccacgcc
     1801 tatcgccgca tgggcttcac cctgtgtggc ctggataccg ctctgtacga tggcaccgcc
     1861 agtgatggcg aacaggccct gtacatgagc atgccctgcc cctaaggccg gccagcccgc
     1921 ctaatgagcg ggcttttttt tccaacacct accacctgca tttgcaacca ggcgaagaga
     1981 ttgtggccaa agcgggcggt ctgcaccgct tcatgaattg gtcggggcca atgctgactg
     2041 attccggtgg ctttcaggtc ttcagcctga gtgaactgcg caagattgaa gagcgcggcg
     2101 tcacctttcg atcgccccgg gatggcgcgg tgattgagtt cacccccgag cgatcgattc
     2161 gcattcaaaa tgcactgggg gccgatgtga tcatggcctt cgatgaatgc ccgccctatc
     2221 cagctgagcg caaggacgtc gaagcggcgg tggcacggac ctatcgctgg ctggagcgct
     2281 gtatcaacgc tcatgaaaga ccccaggatc aagccctatt cggcattgtt caaggcggcg
     2341 tctatctgga tttacgccag caggcggccc gggatctcgt tcagcttgac ctaccgggct
     2401 atgcgatcgg tggtgtcagt gtcggcgaac cctcagaaga aattcaccgc attgtggaag
     2461 cgacagcacc gctcttgccc gcccataaac ctcgctactt gatgggtgtg gggacgtacc
     2521 gcgaaatggt gcaggcgatc gccgctggga ttgacctgtt tgactgcgtg atcccaactc
     2581 gtttggcgcg gcatggtgct gccttagtca agggcgatcg ctggaacctg aaaaatgcac
     2641 agttccgcga agatttccag ccgctggatg aagactgtaa ctgctactgc tgccaaaact
     2701 tcagccgcgc ttacctcaac cacctgattc gatcgcggga aattctgggc tataccttgc
     2761 tctcgattca caacatcacc gagctggtgc gcttcacgac tcggattcgc gaggctatct
     2821 tgagcgatcg cttcgtcgaa gagtttggcc attggttaca gccatccgca gtgcccgtat
     2881 cgccgtgata gcattcagaa caactgagtc gagtcttgcg ttcagagaag ggagttcaac
     2941 gatgtctacc tcctccgctc aatacagtcc tgatttagtc cgtgcttacc tgcaagaaat
     3001 tggccgggtg cggctgctga ccgctgaaga agagttgtgc tttggccgac aggttcagcg
     3061 cctgatgatg ctcttggatg ctcaaaccga gctgcgcgat cgcttggggc atgagcctag
     3121 caaagaagaa tgggctgctg ctgttgacct gaatctcgaa gatctcgatc gccaaattga
     3181 gcaaggtcag cgggcgaaac gcaaaatgat tgaggccaac ctgcgcctcg ttgtttcgat
     3241 cgccaagaaa taccagaagc gtcacatgga attcttggat ctcatccaag aaggcacact
     3301 ggggctggag cgtggtgtcg aaaaattcga tccctccaaa ggctacaagt tctctaccta
     3361 tgcctactgg tggattcgcc aagctattac ccgtgcgatc gcccaacaat cccgcacgat
     3421 tcgtctgcct attcacatca ctgaaaagct gaacaagctc aaaaaaaccc agcgcgaact
     3481 ctcgcagcaa ttgggccgca gtgccacagc ttcagagctg gcagaagtgc tggagctgcc
     3541 actggagcag gtgcgggaat acatccaaat gaaccgccag ccagtatcgc tcgatgtcaa
     3601 agtaggtgac agccaagaca ctgaattgca ggaactgctg gaagacgagc agtcttcgcc
     3661 ctcagattac gtggaacaag aatcgctacg ccgcgatcta cgcaatttaa tggcagaact
     3721 gaccccccag caacaggccg tgattgccct gcgctacggc ttggatgaag gtgacagtct
     3781 gtcgttggcc aaagtcggcg aacgcctgaa tatcagccgc gaagtaccga gctcgaattc
     3841 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3901 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3961 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4021 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4081 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4141 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4201 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4261 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4321 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4381 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4441 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4501 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4561 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4621 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4681 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4741 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4801 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4861 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4921 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4981 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5041 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5101 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5161 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5221 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5281 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5341 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5401 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5461 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5521 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5581 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5641 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5701 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5761 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5821 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5881 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5941 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6001 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6061 ccctttcgtc tcg
//
