LOCUS       sequence-182944-        4866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Renilla Luciferase C-term tagging vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4866)
  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 4866)
  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: pFS346
FEATURES             Location/Qualifiers
     source          1..4866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..31
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(69..91)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     191..210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        291..309
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     291..308
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             370..1302
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="TSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     terminator      1323..1510
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1378..1402)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1585..2704
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1585..1928
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1929..2501
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2507..2704
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2810..2829)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2811..2829)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2916..2933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3087..3675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3167..3186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3846..4637,4638..4706))
                     /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     4469..4488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4707..4811)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
       61 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
      121 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
      181 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg
      241 ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga
      301 cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg gatccccggg
      361 ttaattaaca cttcgaaagt ttatgatcca gaacaaagga aacggatgat aactggtccg
      421 cagtggtggg ccagatgtaa acaaatgaat gttcttgatt catttattaa ttattatgat
      481 tcagaaaaac atgcagaaaa tgctgttatt tttttacatg gtaacgcggc ctcttcttat
      541 ttatggcgac atgttgtgcc acatattgag ccagtagcgc ggtgtattat accagacctt
      601 attggtatgg gcaaatcagg caaatctggt aatggttctt ataggttact tgatcattac
      661 aaatatctta ctgcatggtt tgaacttctt aatttaccaa agaagatcat ttttgtcggc
      721 catgattggg gtgcttgttt ggcatttcat tatagctatg agcatcaaga taagatcaaa
      781 gcaatagttc acgctgaaag tgtagtagat gtgattgaat catgggatga atggcctgat
      841 attgaagaag atattgcgtt gatcaaatct gaagaaggag aaaaaatggt tttggagaat
      901 aacttcttcg tggaaaccat gttgccatca aaaatcatga gaaagttaga accagaagaa
      961 tttgcagcat atcttgaacc attcaaagag aaaggtgaag ttcgtcgtcc aacattatca
     1021 tggcctcgtg aaatcccgtt agtaaaaggt ggtaaacctg acgttgtaca aattgttagg
     1081 aattataatg cttatctacg tgcaagtgat gatttaccaa aaatgtttat tgaatcggac
     1141 ccaggattct tttccaatgc tattgttgaa ggtgccaaga agtttcctaa tactgaattt
     1201 gtcaaagtaa aaggtcttca tttttcgcaa gaagatgcac ctgatgaaat gggaaaatat
     1261 atcaaatcgt tcgttgagcg agttctcaaa aatgaacaat agggcgcgcc acttctaaat
     1321 aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg
     1381 tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact
     1441 ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca
     1501 cctctaccgg cagatccgct agggataaca gggtaatata gatctgttta gcttgcctcg
     1561 tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt
     1621 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt atatttagcc cacacatccc
     1681 catgtataat catttgcatc catacatttt gatggctgca cggcgcgaag caaaaattac
     1741 ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt
     1801 ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc
     1861 tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat
     1921 aaacaaccat gggtaccact cttgacgaca cggcttaccg gtaccgcacc agtgtcccgg
     1981 gggacgccga ggccatcgag gcactggatg ggtccttcac caccgacacc gtcttccgcg
     2041 tcaccgccac cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca
     2101 aggtgttccc cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgacccgg
     2161 actcccggac gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct
     2221 cgtactccgg ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc
     2281 gggggcacgg ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagcggg
     2341 gcgccgggca cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc
     2401 ggcggatggg gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg
     2461 acggcgagca ggcgctctac atgagcatgc cctgccccta atcagtactg acaataaaaa
     2521 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
     2581 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
     2641 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
     2701 actgctgtcg attcgatact aacgccgcca tccagtgttt aaacgagctc gaattcatcg
     2761 atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga
     2821 gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg
     2881 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2941 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3001 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3061 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3601 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3661 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3721 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3781 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3841 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3901 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3961 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4021 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4081 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4141 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4201 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4261 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4321 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4381 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4441 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4501 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4561 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4621 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4681 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4741 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4801 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4861 atgaca
//
