LOCUS       sequence-317199-        6700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Excess Capsid Protein in vivo mRNA display.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6700)
  AUTHORS   Oikonomou P, Salatino R, Tavazoie S
  TITLE     In vivo mRNA display enables large-scale proteomics by next 
            generation sequencing.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Oct 9. pii: 2002650117. doi: 
            10.1073/pnas.2002650117.
  PUBMED    33037152
REFERENCE   2  (bases 1 to 6700)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2020 Oct 9. pii: 2002650117. doi: 
            10.1073/pnas.2002650117."
COMMENT     Sequence Label: plIVMD118
FEATURES             Location/Qualifiers
     source          1..6700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..350
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     regulatory      738..747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             744..1451
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(889..907)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1157..1176
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1470..1493
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1627..1874
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1643..1661)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1892..1911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1893..1911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1921..1938)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1921..1937)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1930..1952)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2078..2533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2375..2396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2664..3467)
                     /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(3468..3683)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3707..3726)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3826..3848
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3886..3904)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3945..4448
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4485..4589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4590..4658,4659..5450)
                     /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(4808..4827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5621..6209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6110..6129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6363..6380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6497..6518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6533..6538,6539..6556,6557..6563)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6571..6587
                     /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     6576..6598
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6595..6611
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6595..6611
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6630..6650
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6632..6650
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttattttttg ctttttctct tgaggtcaca tgatcgcaaa atggcaaatg gcacgtgaag
       61 ctgtcgatat tggggaactg tggtggttgg caaatgacta attaagttag tcaaggcgcc
      121 atcctcatga aaactgtgta acataataac cgaagtgtcg aaaaggtggc accttgtcca
      181 attgaacacg ctcgatgaaa aaaataagat atatataagg ttaagtaaag cgtctgttag
      241 aaaggaagtt tttccttttt cttgctctct tgtcttttca tctactattt ccttcgtgta
      301 atacagggtc gtcagataca tagatacaat tctattaccc ccatccatac tctagaatgc
      361 tagccgttaa aatggcttct aactttactc agttcgttct cgtcgacaat ggcggaactg
      421 gcgacgtgac tgtcgcccca agcaacttcg ctaacgggat cgctgaatgg atcagctcta
      481 actcgcgttc acaggcttac aaagtaacct gtagcgttcg tcagagctct gcgcagaatc
      541 gcaaatacac catcaaagtc gaggtgccta aaggcgcctg gcgttcgtac ttaaatatgg
      601 aactaaccat tccaattttc gccacgaatt ccgactgcga gcttattgtt aaggcaatgc
      661 aaggtctcct aaaagatgga aacccgattc cctcagcaat cgcagcaaac tccggcatct
      721 acgcggatcc accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca
      781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1441 agctgtacaa gtcaagaggg cccttcgaag attataaaga cgatgacgat aagtaaatcg
     1501 ataccgtcga cgatcctacg gtacttattg ccaagaaagc acgagcatca gccgtgcctc
     1561 caggtcgaat cttcaaacga cgacgatcac gcgtcgctcc agtattccag ggttcatcag
     1621 ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     1681 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
     1741 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
     1801 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
     1861 aggctttaat ttgcggccgg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     1921 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     1981 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2041 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     2101 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2161 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2221 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2281 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2341 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2401 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2461 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2521 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2581 gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     2641 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     2701 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     2761 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     2821 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     2881 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     2941 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     3001 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     3061 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     3121 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     3181 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     3241 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     3301 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     3361 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     3421 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     3481 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     3541 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     3601 ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa
     3661 taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg
     3721 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3781 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3841 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3901 tatttttata ggttaatgtc atgataataa tggtttctta gacggatcgc ttgcctgtaa
     3961 cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat
     4021 ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg
     4081 gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca
     4141 tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt
     4201 aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt
     4261 cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc
     4321 cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta
     4381 ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt atttttatag
     4441 cacgtgatga aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4501 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4561 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4621 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4681 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4741 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4801 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4861 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4921 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4981 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5041 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5101 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5161 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5221 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5281 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5341 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5401 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5461 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5521 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5581 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5641 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5701 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5761 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6181 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6241 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6301 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6361 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     6421 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     6481 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     6541 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     6601 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     6661 tggagctccg gatgcaaggg ttcgaatccc ttagctctca
//
