LOCUS       sequence-232099-        9746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SigF2 epitope tagged construct for integration at neutral site 2.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9746)
  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 9746)
  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: KFP08
FEATURES             Location/Qualifiers
     source          1..9746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     333..352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(571..675)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     743..761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(788..793,794..810,811..816)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             3648..3713
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      complement(4182..4209)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4301..4387)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(4761..4766,4767..4783,4784..4789)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4978..5511
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(6601..6620)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             8078..8269
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(8263..8285)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    8371..8511
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     8426..8445
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8526..8543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8697..9285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8777..8796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9456..9746,1..501,502..570))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
       61 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      301 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
      361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
      541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
      601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
      721 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt
      781 ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt
      841 gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca
      901 ccgtcaccct ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc
      961 gggatatcgt ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg
     1021 cgttgatgca atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc
     1081 gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca
     1141 cacccgtcct gtggatccca ggctcagtgt ggttcgcatc ttttgatgat caccactgga
     1201 cgcaacacag tagggcaatt caagagcatc caaagcctct tcaaccccag gaacggcttg
     1261 tagatgcgtt tctaacgcga tacgggtacg gcgttgatag tgctgaacaa agtcaggggg
     1321 tggaggattg cctaaccgtc gctcaattag tttgagacag tcagccatgg aatgacccac
     1381 aaactgctca aacatgtcat ccaaagtcac caacagaccc agttcattga gcatgtctgc
     1441 aaagacgcga ttagtgatgc gttcgctatc aacaagcaca ccatcacagt cgaaaatcac
     1501 cagctgaaac ggtgaagttt gcattgtttt taagcacgag ccatcaacag taagagcgat
     1561 cgcgctggga cgatgtaatc gcgccgtagg cagggttttg cctcatccgg cagatgacaa
     1621 gcttcaagat tcggcagtga agtatcgagc aagcgatacc aaacgcggcc gcgaggaggc
     1681 ctcggcaact ggaagcgcag gtcttcccag taagcattaa aggctaggta aagccattcc
     1741 tgctggcgag gatggcagag actgacggcc agactgtggg accacagcgc ccaatcgggt
     1801 tgtttgagtt tgacgccatg ccagatggca tagggacgac gcggatgggg ttcgttctgc
     1861 agcagttcgt tctgttggaa catcaccagc gactgggaaa gttcaatcag gcggcgactg
     1921 aacaccaaga aatcggcatg gcgatcgcac agcgaccaat caaaccagct gatctcattg
     1981 tcttggcagt aggcgttatt gttaccctgc tgactgcgtt tgacctcatc gcccatcgtc
     2041 agcatcggtg tgccctgggc gaggaataac gtggcgagca aattgcgctg ctgccgttcc
     2101 cgtaagctca gaatcgtggg gtcatcggtc tctccttcaa tgccgtagtt ccagctgtag
     2161 ttgtcattgg tcccgtcccg attgttctct ccattggcaa agttgtgctt ctggctatag
     2221 ctgactcgtc acgggtaacc gatatcgtcg acaggcctgc aacttgctgg ttattcccat
     2281 tgaccaagcg ctgctctacg ttgagccact ctacttagaa gcctcgcgca atagcttgcc
     2341 agcgctcacc cgcgtcatta cggcctacca agatcgcatt gtgatgactc ccagcctcct
     2401 cgaaagcctg caaaaactgt ttccagactc aacacccgcc ctgacgccgc ttgaacaacc
     2461 ggtgttgacg accgagcagt ccgctgtcct caaccctgac cagccctaga aacccatcgt
     2521 tttccagatt ggcgttgcca atgggcgggc tgaactagca gaactcaatg gtcagcggat
     2581 caggtgattg atcaaggggc gttgaccttg gtcatgctca aagtgccgca acatttcttc
     2641 attgttgcga gagcatgttc tgattaaccg aagatcgcga taacggccca aaatttcagc
     2701 tttagcaacg cttttgagtc acgaatatcg agggagtatc aaggattatt gcagttcata
     2761 gcactcctct agtccccttc gtacggtggg aaaagagagc gacagcaaac tccctgttta
     2821 ggatgcccaa gttctctttg cctcacacag tcctcggagg cctgatagtg gtgaaccaag
     2881 cagcgttacg aacccgaggg atggaattgc tggtcgccta cgctgagact cccagcattg
     2941 ccctccgcaa ccagattgtt cagctcaatc tagggctggt ccgcaaaatt gcccaccgat
     3001 taagccggca atgctctgaa ccctatgaag atctggaaca gattggtttc atgggattaa
     3061 ttcgagcgat cgagcgattt cagcctcatc aagggtgtgc atttagttcc tttgcagtgc
     3121 cctatatacg cggcgaaatt cttcattttt tacgcgacag ttgtcatagc attcggattc
     3181 cgcgccgctg gcaagaactg cagcgtcaag gacagcaatt gcgggaggtc ttgcgccatg
     3241 aatggggacg agccccaacg gaacaagagc tggcttttgc tctcaatctg tcaatgacgg
     3301 aatggcaaga aattcacctc gctcgccgta atcggatgcc gatgagtctg gatgccacgc
     3361 tcagtcaagc cggtgattct tccgtcacct tggtggaaac cctgcccgat accagccagc
     3421 agtggcagca ggccgccctt gaagagcgtc aacagctggt ggaagcgatc ggccaactgg
     3481 aagaaaaagc cagaacagcg atcgaatctg tttacctgcg cgggctaacc cgcaaagaag
     3541 cggcccagca atttggtatc agtccgatga cgattacccg ccggatcaat cgcggggttc
     3601 aaaaaatcgc cgagcacctc gaccgcatgg caaacgagcc cttagccgac tacaaggacc
     3661 acgacggtga ctacaaggac cacgacatcg actacaagga cgacgacgac aagtgagtcg
     3721 cttccctgtc cagtcctcct gtctgtctta agcagcttag gcgcgatcgc ctaggctgtt
     3781 tttttgaccg tccattgcgt gatcggtgcc agcgatttcc agccgagaaa gcggccgatc
     3841 ggttccgctc gctgcaccgc cacacggatt aaatctcccc cccagcgctg ctgtccagcc
     3901 aaaacgactt gctcgctttc gagggtaacg gcgttaacaa ctagccgccc acccatcggc
     3961 agcgcttgcc agcattgatc gagcacacct ggagcagtga caccgccacc gatgaagatg
     4021 gcatccggtt ggggcaagcc actcagcgta gccggtgctg aaccggcaac cacttgcagg
     4081 ttcggaaccc cgagggcgat cgcattctgt gtaatgatta cccgggctcg agctagcaag
     4141 cttggccgga tccggccgga tccggagttt gtagaaacgc aaaaaggcca tccgtcagga
     4201 tggccttctg cttaatttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     4261 cgggccgttg cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     4321 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     4381 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg
     4441 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
     4501 ccaggcaaat tctgttttat cagccgttac cccacctact agctaatccc atctgggcac
     4561 atccgatggc aagaggcccg aaggtccccc tctttggtct tgcgacgtta tgcggtatta
     4621 gctaccgttt ccagtagtta tccccctcca tcaggcagtt tcccagacat tactcacccg
     4681 tccgccactc gtcagcaaag aagcaagctt agatccacta gtgacgcaca ccgtggaaac
     4741 ggatgaaggc acgaacccag ttgacataag cctgttcggt tcgtaaactg taatgcaagt
     4801 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     4861 cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg cctcgggcat
     4921 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     4981 ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc
     5041 tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa atccatgcgg
     5101 gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc ccaacatcag
     5161 ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc
     5221 ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccaa gtttgagcag
     5281 ccgcgtagtg agatctatat ctatgatctc gcagtctccg gcgagcaccg gaggcagggc
     5341 attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg tgcttatgtg
     5401 atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac aaagttgggc
     5461 atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta acaattcgtt
     5521 caagccgaga tcggcttccc ggccgcggag ttgttcggta aattgtcaca acgccgcact
     5581 agtggatctg gtaaccccag cgcggagatc tcgcagcgta aagccgtcat ggcaggtgat
     5641 gaagttaatg gtgcgtccgg cataccattg gtctgtgctg tagacatcgg ggctacccag
     5701 caggcgttga ctgagggcgt aagtacagcc ctgatctcca cgccaaaaac gccgaatatc
     5761 gtcccggaag ggaccgttcc aagtcccaaa gcgatcgcca ataaaggtac caacctgata
     5821 taagccggct gcgtcccaag cttcagcaat gagcttcgta ccggccaaaa ccggatcgga
     5881 atcaatcgcc caaagcaagg gcggatccga tagggggttg ccattggcat cacgactcag
     5941 caccgacgca aggtcaaagc ggaagccatc gacgtgcatt tccgagaccc aataacgcag
     6001 gcaatcgaga atcagcgatc gcacgatcga attgttagct ttgacggtgt tgccgcagcc
     6061 ggtgtagttg cgatagcccg cctgttcatc cagcagatag taggttgaat tcgctagacc
     6121 tttgaaagac agcgttggac cgtcttcatt cccttcagca gtgtggttga acaccacgtc
     6181 gagaatcacc tcaatccctg cttggtggag cgccttgacg aggtcgcgga actcatcaac
     6241 tggaccaagt ggatcatggc gagagctgta ggctgcgtgg ggcgcaaaaa aagccatggt
     6301 gctgtagccc cagtagttct cgcgtcctaa gggggcatct tggcgatcga actggtgcac
     6361 cggcagcaac tcaacggccg tgacgccgag ggattgcagg tagggaattt tttcgattag
     6421 accagcgtag gtgccacgtt tttcaggggc gacgccggaa ttgggatgct tggtgaagcc
     6481 gccaacatgc agctcataga ttactgtgcg agcgtagggt gtgcgtggat ggcgatcgga
     6541 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     6601 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     6661 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     6721 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
     6781 gcttcctaat gcaggagtcg cataagggag agcgtcgacc gatgcccttg agagccttca
     6841 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     6901 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     6961 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     7021 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     7081 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     7141 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     7201 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     7261 gatcgctcgc ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga
     7321 tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat
     7381 accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa
     7441 tggaagccgg cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt
     7501 cttgcggaga actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc
     7561 catctccagc agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg
     7621 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     7681 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     7741 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     7801 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
     7861 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
     7921 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
     7981 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     8041 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     8101 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     8161 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     8221 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     8281 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     8341 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     8401 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     8461 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     8521 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     8581 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     8641 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     8701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     8761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     9001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     9061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     9121 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     9181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     9241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     9301 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     9361 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     9421 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     9481 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     9541 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     9601 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     9661 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     9721 agagtaagta gttcgccagt taatag
//
