LOCUS       sequence-182945-        5818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Beetle Luciferase C-term tagging vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  AUTHORS   Keifenheim D, Sun XM, D'Souza E, Ohira MJ, Magner M, Mayhew MB, 
            Marguerat S, Rhind N
  TITLE     Size-Dependent Expression of the Mitotic Activator Cdc25 Suggests a 
            Mechanism of Size Control in Fission Yeast.
  JOURNAL   Curr Biol. 2017 May 22;27(10):1491-1497.e4. doi: 
            10.1016/j.cub.2017.04.016. Epub 2017 May 4.
  PUBMED    28479325
REFERENCE   2  (bases 1 to 5818)
  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.cub.2017.04.016"; journalName: "Curr Biol"; date: 
            "2017-05-22- 22"; volume: "27"; issue: "10"; pages: "1491-1497.e4"
COMMENT     Sequence Label: pFS345
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..23
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(61..83)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     183..202
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        283..301
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     283..300
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             362..2011
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(425..443)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      2032..2219
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2087..2111)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2294..3650
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2294..2637
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2638..3447
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2705..2724)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3453..3650
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3754..3773)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3755..3773)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3860..3877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4031..4619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4111..4130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4790..5581,5582..5650))
                     /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     5413..5432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5651..5755)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
       61 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      121 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      181 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
      241 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
      301 aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga cggatccccg ggttaattaa
      361 cgaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg aagatggaac
      421 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
      481 ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg aaatgtccgt
      541 tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg
      601 cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc
      661 agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgggcatttc
      721 gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa
      781 aaagctccca atcatccaaa aaattattat catggattct aaaacggatt accagggatt
      841 tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt
      901 tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact cctctggatc
      961 tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga gattctcgca
     1021 tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt
     1081 tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg
     1141 agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc aggattacaa
     1201 gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa gcactctgat
     1261 tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc ccctctctaa
     1321 ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg
     1381 gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc
     1441 ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac
     1501 gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta tgtccggtta
     1561 tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg
     1621 agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc tgaagtctct
     1681 gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct tgctccaaca
     1741 ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc
     1801 cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta
     1861 cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga
     1921 agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa
     1981 ggccaagaag ggcggaaaga tcgccgtgta gggcgcgcca cttctaaata agcgaatttc
     2041 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     2101 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     2161 gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc
     2221 agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg
     2281 tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
     2341 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
     2401 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
     2461 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
     2521 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
     2581 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
     2641 ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga tgctgattta
     2701 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     2761 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2821 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2881 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
     2941 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     3001 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     3061 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     3121 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     3181 aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     3241 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     3301 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     3361 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     3421 catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt cttgttttca
     3481 agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta
     3541 gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc
     3601 agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc
     3661 gatactaacg ccgccatcca gtttaaacga gctcgaattc atcgatgata tcagatccac
     3721 tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga
     3781 taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3841 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3901 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3961 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4021 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4081 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4141 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4201 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4261 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4321 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4381 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4441 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4501 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4561 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4621 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4681 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4741 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4801 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4861 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4921 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4981 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5041 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5101 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5161 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5221 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5281 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5341 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5401 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5461 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5521 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5581 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5641 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5701 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5761 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaaccta
//
