LOCUS       sequence-237903-        5521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sigF2∆::NatR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5521)
  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 5521)
  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: KFP78
FEATURES             Location/Qualifiers
     source          1..5521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3539..3556)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3539..3555)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3548..3570)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3764..3783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3883..3905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3943..3961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4029..4133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4134..4202,4203..4994)
                     /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(4352..4371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5165..5521,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaattcga gctcggtacc
      661 ctgctaccga tttctttgcg atcgctggtt ggctaggctg ggcaaccctc gccctgacga
      721 ttttgactgt gctggcgttt gtccttgggt ggggctttcg cttccgactg gtgggggtga
      781 ccgccttcac gatcgtgctc attggcggct gcattggctt tgccctcggc ttcaagcagc
      841 ctgtcagcat tgaaggtgct gctcctttta acgtggtcta cgacgctggg ggcagtgagg
      901 ccgtggttgc cgttgccccg acgatcgcgc cagaagcggt tgagccgacc ctgaagcagg
      961 cagccttcaa cctcttttca cgtggccgct acagcagcca gggcgaacaa cagctccata
     1021 tccgcctgcg tacgattctt catccccagc ccggaatctc tgagccgctc tacctcggcg
     1081 aggtgaagcg atcgctgaat caactcgatg acgaggcgat gcaaatcctt gtcgatcccc
     1141 aggcgattgc tcgcttggcc gctgccgaaa atggctgagt cggtgccggc tcaggggtgg
     1201 ccgctgacgc agctcccagg gctgagcaat ggcgatcgcc aacaactggc agcgcacggc
     1261 attcacttca gcctggactt actcaagctg cgatcggaag cggcgcaact ggacttagct
     1321 cagcagctgc gacagcccgt ccagcgggtg cgcaaatggc aagctttggc acagctttcg
     1381 catctgccct cggtcaacca tcgttggtgt ggcgtgctgc tgcacacggg catcagtagc
     1441 ccgcaaatgc tggcgcttca gagtccgccc caattgcatc aacgcctgca tcgtctccag
     1501 cgatcgctat tgggcgatcg cagtccagcc ccgacccttg ggcaagtcga tcgctggatt
     1561 cgcgaagccc aacagttttt gcaacggcgc taggctaagg gctcgtttgc catgcggtcg
     1621 aggtgctcgg cgattttttg aaccccgcga ttgatccggc gggtaatcgt caaaaaaaaa
     1681 gcccgctcat taggcgggct ggccggcctt aggggcaggg catgctcatg tacagggcct
     1741 gttcgccatc actggcggtg ccatcgtaca gagcggtatc caggccacac agggtgaagc
     1801 ccatgcggcg ataggcgtgg atagcgggag cgttcacgtt ggtcacttcc agccacaggt
     1861 ggccagcacc gcgttcgcga gcaaattcgg tggccaggcc catcagagcg cgaccaacgc
     1921 cgtgaccgcg gtgttcggga gccacttcaa tatcttccac ggtcaagcgg cgattccagc
     1981 cgctgtagct caccaccaca aagccagcca gatcgccatc atcaccgtag gccacaaagg
     2041 tgcggctatc gggatcgcca tcttcgccag catccgattc atcatcactt tcatcatcgg
     2101 gaaacacttt ggtcagaggg ggatcgacgg gcacttcgcg cagggtaaag ccatcacccg
     2161 tggcggtcac gcgaaacacg gtatcggtgg taaagctgcc atccagggct tcgatggctt
     2221 cggcatcacc gggaacgctg gtgcggtagc ggtaggcggt atcatccagg gtggtcatat
     2281 ggatccggcc accgtttgct tttaacgtca gccttttacc taaattccca tcaagcgatc
     2341 ttttaatatc ggacaacgtc atcgccattg catccacctc actacattta ttgtacaaca
     2401 cgagcccatt tttgtcaaat aaaatttaaa ttatatcaac gttaataaga cgttgtcaat
     2461 aaaattattt tgacaaaatt tcctttgctt gtttgctcta ttcctataat tcgatgataa
     2521 gctgtcaaac atgagaattg cctccgagga ctgtgtgagg caaagagaac ttgggcatcc
     2581 taaacaggga gtttgctgtc gctctctttt cccaccgtac gaaggggact agaggagtgc
     2641 tatgaactgc aataatcctt gatactccct cgatattcgt gactcaaaag cgttgctaaa
     2701 gctgaaattt tgggccgtta tcgcgatctt cggttaatca gaacatgctc tcgcaacaat
     2761 gaagaaatgt tgcggcactt tgagcatgac caaggtcaac gccccttgat caatcacctg
     2821 atccgctgac cattgagttc tgctagttca gcccgcccat tggcaacgcc aatctggaaa
     2881 acgatgggtt tctagggctg gtcagggttg aggacagcgg actgctcggt cgtcaacacc
     2941 ggttgttcaa gcggcgtcag ggcgggtgtt gagtctggaa acagtttttg caggctttcg
     3001 aggaggctgg gagtcatcac aatgcgatct tggtaggccg taatgacgcg ggtgagcgct
     3061 ggcaagctat tgcgcgaggc ttctaagtag agtggctcaa cgtagagcag cgcttggtca
     3121 atgggaataa ccagcaagtt gccttcggca gcgcgggagc cttcgcggtt ccagagcgta
     3181 atttgctcag aaatctgagg atcttggttg atgcgagccg tgatttgctc gggaccaaaa
     3241 atgaggcgct ggcgcggaaa ttcgtagagc aagaggcgac cgtattcctc gccgtctgat
     3301 cgccccgcca gccaagcaat catattgtta cgccccaatg gtgtgaaggg cgtcaggatc
     3361 aggaactctt cgctcgcagc ggtcggtaag cgcgtaatta agtagtaggg ctgaacggtt
     3421 gccgtctcgc cagcgtagat ttcgcgtggg taggcccact ggtcatcgcg gttgtagaaa
     3481 acctgtggat ggtacccggg gatcctctag agtcgacctg caggcatgca agcttggcac
     3541 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3601 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3661 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     3721 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3781 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3841 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3901 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3961 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4141 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4201 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4261 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4321 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4381 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4441 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4501 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4561 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4621 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4681 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4741 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4801 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4861 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4921 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4981 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5521 c
//
