LOCUS       sequence-209214-        7634 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 7634)
  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 7634)
  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..7634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1961..1972
                     /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(2803..2819)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2803..2819)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2851..2870)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2852..2870)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2877..2894)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2877..2893)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2886..2908)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3034..3489
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3121..3140)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3331..3352
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3620..4423)
                     /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"
     promoter        complement(4424..4639)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4663..4682)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4782..4804
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4842..4860)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4901..5404
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5441..5545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5546..5614,5615..6406)
                     /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     complement(5764..5783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6577..7165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7066..7085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7319..7336
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7453..7474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7489..7494,7495..7512,7513..7519)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7527..7543
                     /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)."
     primer_bind     7532..7554
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7551..7567
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7551..7567
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7588..7606
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     join(7634,1..16)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cgaggtcgac ggtatcgata ataaacatat taaagctttc aaaaatatag tctttcgaag
       61 agctttttca tactaagaga agcgttgcta atattccgaa ttaatcacgc aattgcctct
      121 ctgctttcca cgtaacccat cataggaaat tgtattaata tgttacttac tggaattgca
      181 caggaaaaca ccgacaaaaa attggtttgt ctgtgttttg acggagagct actgacttgt
      241 aggttcactt tttcacacac cgcactatgc atatatgttg catttttatt attcatccgc
      301 acgtggctgc acacgtgaat gaaaaagaag caatactatc tttatatatt gaggcaatca
      361 gaaagaagtt acaggctaac aatcaaatcg gccaataaaa gagcataaca aggcaggaac
      421 agctatgaag tttggtgagc acttgagcaa gtctcttatt agacagtaca gctactatta
      481 tattagttat gatgatctga agactgagct agaagataac ttatctaaga ataacggtca
      541 gtggacgcaa gaattggaaa cagatttttt agaatctttg gagatcgagt tagacaaggt
      601 ttacacattt tgcaaagtta agcatagtga agtttttaga cgtgtaaaag aggttcaaga
      661 acaagtgcaa catactgttc gtttgttaga ctccaataat cctcctactc agctagattt
      721 tgagattcta gaagaagaac taagtgatat cattgccgat gttcatgatc tagcaaagtt
      781 ttcgagacta aactacactg gtttccaaaa gattatcaag aaacacgata agaagacagg
      841 ttttatcttg aaaccagttt tccaagttag attagactct aaaccatttt tcaaggaaaa
      901 ctatgacgaa ctagtcgtta aaatttccca actatatgat attgccagaa cttcaggtcg
      961 cccaatcaag ggagactcat ctgctggtgg gaagcaacaa aatttcgtta gacagacaac
     1021 aaagtattgg gttcaccctg ataacattac agaattgaag ctgatcatct tgaagcattt
     1081 accagtgtta gtcttcaaca ctaataagga attcgaaaga gaagattccg ctattacttc
     1141 gatctatttt gataatgaaa atctagacct ttattatggc agattaagaa aagatgaagg
     1201 tgcagaagcc cacagattga gatggtatgg gggtatgtct acagacacga tctttgtaga
     1261 aagaaagacc catagagagg attggactgg tgagaaatct gtcaaggcaa gatttgcgct
     1321 aaaggaacgt catgttaatg acttcttaaa gggtaaatat actgttgatc aagtatttgc
     1381 caagatgcgt aaagaaggca agaagccaat gaacgaaatt gaaaacttgg aggcgttagc
     1441 atccgaaatt caatacgtca tgttaaagaa aaagttgaga cctgtagtga gatcattcta
     1501 caatagaact gcttttcaat tacctggtga tgcaagagtt cgtatctccc ttgatacaga
     1561 gttaactatg gtgagagaag acaactttga tggtgtggat agaactcata agaattggag
     1621 gagaactgat attggtgtcg attggccatt caagcagcta gatgacaagg atatttgccg
     1681 tttcccatat gcagtcctgg aagttaaatt gcaaactcaa ctaggtcaag aacctccaga
     1741 gtgggtacgt gaattagtcg ggtctcactt agttgagcca gtcccaaaat tctccaagtt
     1801 tattcatggt gtggctacct tactaaatga taaagtggat tcaatcccat tttggttacc
     1861 tcaaatggat gttgatatca ggaaacctcc actaccaaca aatattgaaa taacaagacc
     1921 cggtagatcc gataacgaag ataacgactt cgatgaggat gatgaggatg acgctgcttt
     1981 ggttgctgcc atgacaaacg cacccggtaa ttctcttgat atagaagaat ctgttggtta
     2041 cggggctact tcggccccca cctccaacac taatcatgtt gttgagagtg ctaatgctgc
     2101 ttattatcaa agaaagatca ggaatgccga aaatcctatc tccaagaaat actatgaaat
     2161 cgtggcattt tttgaccact attttaacgg tgatcaaata tctaaaatcc caaaaggtac
     2221 tacttttgat actcaaattc gtgccccacc gggaaagacg atatgtgtgc cagttcgtgt
     2281 ggaaccaaaa gtttactttg ccactgaaag aacctacctg tcttggttgt ccatctcgat
     2341 attgttgggc ggtgtctcca ctactttatt aacatatggt tcaccaacag ccatgattgg
     2401 ctccattggg ttctttatta cttccttggc tgtgttgata cgtactgtta tggtttatgc
     2461 caagagagtg gtaaacatta gactaaagag agcagtagat tatgaggata agatcgggcc
     2521 cggcatggtt tctgtgtttc taattctctc tattttattt tcctttttct gcaacctagt
     2581 tgctaaatga atacacagcg tgtttttttt actgtataat taagtaataa tcatattaga
     2641 tataataatt tctttagcgc cattttttta tatatgagaa ctaccctctt gcctgaaatg
     2701 cacaccttca ccaggcgcac actggttcaa aaatagtaaa caaaatcatg cgaaaataca
     2761 ataaaagacg ttaacttgat agaatgaaac tgcagcccgg gggatccact agttctagag
     2821 cggccgccac cgcggtggag ctccaattcg ccctatagtg agtcgtatta caattcactg
     2881 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2941 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3001 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     3061 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3121 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3181 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3241 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3301 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3361 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3421 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3481 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3541 agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat
     3601 ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc
     3661 ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca aatagtcctc
     3721 ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt ctatactgtt
     3781 gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt
     3841 aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt
     3901 cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc
     3961 atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct
     4021 gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta atgtctgccc
     4081 attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag
     4141 caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct
     4201 taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt agtaaacaaa
     4261 ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg
     4321 aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac
     4381 taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc
     4441 tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa
     4501 cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     4561 gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa
     4621 aaaaaaatga tgaattgaaa aggtggtatg gtgcactctc agtacaatct gctctgatgc
     4681 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4741 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4801 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4861 tttataggtt aatgtcatga taataatggt ttcttagacg gatcgcttgc ctgtaactta
     4921 cacgcgcctc gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat
     4981 ttttatgttt tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat
     5041 gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata
     5101 tatttattag acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa
     5161 tgtaaaatca caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc
     5221 attaatacct gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta
     5281 gagtctttta catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt
     5341 ctatttttaa tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg
     5401 tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5461 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5521 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5581 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5641 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5701 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5761 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5821 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5881 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5941 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6001 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6061 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6121 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6181 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6241 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6301 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6361 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6421 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6481 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6541 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6601 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6661 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6721 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6781 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6841 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6901 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6961 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7021 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7081 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7141 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7201 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7261 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7321 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7381 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     7441 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     7501 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     7561 ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctggt
     7621 accgggcccc ccct
//
