LOCUS       sequence-205003-        7636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VTC4 sequence expressed in yeast plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7636)
  AUTHORS   Bentley-DeSousa A, Holinier C, Moteshareie H, Tseng YC, Kajjo S, 
            Nwosu C, Amodeo GF, Bondy-Chorney E, Sai Y, Rudner A, Golshani A, 
            Davey NE, Downey M
  TITLE     A Screen for Candidate Targets of Lysine Polyphosphorylation 
            Uncovers a Conserved Network Implicated in Ribosome Biogenesis.
  JOURNAL   Cell Rep. 2018 Mar 27;22(13):3427-3439. doi: 
            10.1016/j.celrep.2018.02.104.
  PUBMED    29590613
REFERENCE   2  (bases 1 to 7636)
  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.2018.02"; journalName: "Cell Rep"; date: 
            "2018-03-27- 27"; volume: "22"; issue: "13"; pages: "3427-3439"
COMMENT     Sequence Label: pRS316-VTC4
FEATURES             Location/Qualifiers
     source          1..7636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2017..2033
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2017..2033
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2864..2875
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     primer_bind     complement(4816..4832)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4817..4833)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4862..4880)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4901..4917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4901..4917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4914..4936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4925..4941
                     /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(4949..4955,4956..4973,4974..4979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5132..5149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5303..5891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6062..6853,6854..6922))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6685..6704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6923..7027)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7064..7567
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7609..7627
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg atcccccggg
     2041 ctgcagtttc attctatcaa gttaacgtct tttattgtat tttcgcatga ttttgtttac
     2101 tatttttgaa ccagtgtgcg cctggtgaag gtgtgcattt caggcaagag ggtagttctc
     2161 atatataaaa aaatggcgct aaagaaatta ttatatctaa tatgattatt acttaattat
     2221 acagtaaaaa aaacacgctg tgtattcatt tagcaactag gttgcagaaa aaggaaaata
     2281 aaatagagag aattagaaac acagaaacca tgccgggccc gatcttatcc tcataatcta
     2341 ctgctctctt tagtctaatg tttaccactc tcttggcata aaccataaca gtacgtatca
     2401 acacagccaa ggaagtaata aagaacccaa tggagccaat catggctgtt ggtgaaccat
     2461 atgttaataa agtagtggag acaccgccca acaatatcga gatggacaac caagacaggt
     2521 aggttctttc agtggcaaag taaacttttg gttccacacg aactggcaca catatcgtct
     2581 ttcccggtgg ggcacgaatt tgagtatcaa aagtagtacc ttttgggatt ttagatattt
     2641 gatcaccgtt aaaatagtgg tcaaaaaatg ccacgatttc atagtatttc ttggagatag
     2701 gattttcggc attcctgatc tttctttgat aataagcagc attagcactc tcaacaacat
     2761 gattagtgtt ggaggtgggg gccgaagtag ccccgtaacc aacagattct tctatatcaa
     2821 gagaattacc gggtgcgttt gtcatggcag caaccaaagc agcgtcatcc tcatcatcct
     2881 catcgaagtc gttatcttcg ttatcggatc taccgggtct tgttatttca atatttgttg
     2941 gtagtggagg tttcctgata tcaacatcca tttgaggtaa ccaaaatggg attgaatcca
     3001 ctttatcatt tagtaaggta gccacaccat gaataaactt ggagaatttt gggactggct
     3061 caactaagtg agacccgact aattcacgta cccactctgg aggttcttga cctagttgag
     3121 tttgcaattt aacttccagg actgcatatg ggaaacggca aatatccttg tcatctagct
     3181 gcttgaatgg ccaatcgaca ccaatatcag ttctcctcca attcttatga gttctatcca
     3241 caccatcaaa gttgtcttct ctcaccatag ttaactctgt atcaagggag atacgaactc
     3301 ttgcatcacc aggtaattga aaagcagttc tattgtagaa tgatctcact acaggtctca
     3361 actttttctt taacatgacg tattgaattt cggatgctaa cgcctccaag ttttcaattt
     3421 cgttcattgg cttcttgcct tctttacgca tcttggcaaa tacttgatca acagtatatt
     3481 taccctttaa gaagtcatta acatgacgtt cctttagcgc aaatcttgcc ttgacagatt
     3541 tctcaccagt ccaatcctct ctatgggtct ttctttctac aaagatcgtg tctgtagaca
     3601 tacccccata ccatctcaat ctgtgggctt ctgcaccttc atcttttctt aatctgccat
     3661 aataaaggtc tagattttca ttatcaaaat agatcgaagt aatagcggaa tcttctcttt
     3721 cgaattcctt attagtgttg aagactaaca ctggtaaatg cttcaagatg atcagcttca
     3781 attctgtaat gttatcaggg tgaacccaat actttgttgt ctgtctaacg aaattttgtt
     3841 gcttcccacc agcagatgag tctcccttga ttgggcgacc tgaagttctg gcaatatcat
     3901 atagttggga aattttaacg actagttcgt catagttttc cttgaaaaat ggtttagagt
     3961 ctaatctaac ttggaaaact ggtttcaaga taaaacctgt cttcttatcg tgtttcttga
     4021 taatcttttg gaaaccagtg tagtttagtc tcgaaaactt tgctagatca tgaacatcgg
     4081 caatgatatc acttagttct tcttctagaa tctcaaaatc tagctgagta ggaggattat
     4141 tggagtctaa caaacgaaca gtatgttgca cttgttcttg aacctctttt acacgtctaa
     4201 aaacttcact atgcttaact ttgcaaaatg tgtaaacctt gtctaactcg atctccaaag
     4261 attctaaaaa atctgtttcc aattcttgcg tccactgacc gttattctta gataagttat
     4321 cttctagctc agtcttcaga tcatcataac taatataata gtagctgtac tgtctaataa
     4381 gagacttgct caagtgctca ccaaacttca tagctgttcc tgccttgtta tgctctttta
     4441 ttggccgatt tgattgttag cctgtaactt ctttctgatt gcctcaatat ataaagatag
     4501 tattgcttct ttttcattca cgtgtgcagc cacgtgcgga tgaataataa aaatgcaaca
     4561 tatatgcata gtgcggtgtg tgaaaaagtg aacctacaag tcagtagctc tccgtcaaaa
     4621 cgcagacaaa ccaatttttt gtcggtgttt tcctgtgcaa ttccagtaag taacatatta
     4681 atacaatttc ctatgatggg ttacgtggaa agcagagagg caattgcgtg attaattcgg
     4741 aatattagca acgcttctct tagtatgaaa aagctcttcg aaagactata tttttgaaag
     4801 ctttaatatg tttatcgata ccgtcgacct cgaggggggg cccggtaccc agcttttgtt
     4861 ccctttagtg agggttaatt ccgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4921 gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag
     4981 cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt
     5041 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5101 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5161 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5221 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5281 aaaaggccgc gttgctggcg tttttccata ggctcggccc ccctgacgag catcacaaaa
     5341 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttcc
     5401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5461 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5701 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5821 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5881 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5941 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6001 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6061 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6121 tagttgcctg actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6181 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6241 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6301 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6361 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6421 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag
     6481 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6541 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6601 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6661 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6721 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6781 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6841 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6901 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6961 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7021 ttccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa
     7081 taattataat ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag
     7141 aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc
     7201 aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc
     7261 atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt
     7321 taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta
     7381 cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct
     7441 tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca
     7501 gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca
     7561 agcgatccgt cctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7621 acgaggccct ttcgtc
//
