LOCUS       sequence-238571-        5642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a part of RPS14 protein with a flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5642)
  AUTHORS   Lessard F, Igelmann S, Trahan C, Huot G, Saint-Germain E, Mignacca 
            L, Del Toro N, Lopes-Paciencia S, Le Calve B, Montero M, 
            Deschenes-Simard X, Bury M, Moiseeva O, Rowell MC, Zorca CE, 
            Zenklusen D, Brakier-Gingras L, Bourdeau V, Oeffinger M, Ferbeyre G
  TITLE     Senescence-associated ribosome biogenesis defects contributes to 
            cell cycle arrest through the Rb pathway.
  JOURNAL   Nat Cell Biol. 2018 Jul;20(7):789-799. doi: 
            10.1038/s41556-018-0127-y. Epub 2018 Jun 25.
  PUBMED    29941930
REFERENCE   2  (bases 1 to 5642)
  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: 
            "10.1038/s41556-018-0127-y"; journalName: "Nat Cell Biol"; date: 
            "2018-07"; volume: "20"; issue: "7"; pages: "789-799"
COMMENT     Sequence Label: pcDNA3-FLAG-RPS14(89-151)
FEATURES             Location/Qualifiers
     source          1..5642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             305..328
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(581..599)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(582..599)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(619..636)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    625..849
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      895..1323
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(982..1001)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1192..1213
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1332..1352)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1337..1666
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1517..1652
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1579..1598
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1733..2527
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1787..1806)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2397..2416
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2701..2822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2738..2757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2792..2811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2871..2887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2871..2887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2884..2906)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2895..2911
                     /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(2919..2925,2926..2943,2944..2949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2964..2985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3102..3119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3273..3861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3353..3372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4032..4823,4824..4892))
                     /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     4655..4674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4893..4997)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5072..5091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5263..5642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc tcggatccac
      301 catggattac aaggatgacg atgacaaggg tatcaccgcc ctacacatca aactccgggc
      361 cacaggagga aataggacca agacccctgg acctggggcc cagtcggccc tcagagccct
      421 tgcccgctcg ggtatgaaga tcgggcggat tgaggatgtc acccccatcc cctctgacag
      481 cactcgcagg aaggggggtc gccgtggtcg ccgtctgtga gaattctgca gatatccatc
      541 acactggcgg ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
      601 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
      661 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
      721 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
      781 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
      841 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
      901 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      961 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1021 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1081 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1141 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1201 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1261 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1321 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     1381 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     1441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     1501 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     1561 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     1621 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     1681 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     1741 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1801 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1861 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     1921 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1981 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2041 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2101 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2161 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2221 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     2281 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2341 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2401 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2461 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2521 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     2581 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2641 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2821 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     2881 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2941 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3001 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3061 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3241 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4021 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4081 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4141 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4201 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4261 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4321 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4381 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4441 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4501 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4561 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4621 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4681 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4741 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4801 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4861 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4921 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4981 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     5041 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5101 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     5161 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     5221 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     5281 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     5341 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     5401 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     5461 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     5521 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     5581 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     5641 tg
//
