LOCUS       sequence-317197-        6594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Excess Capsid Protein for in vivo mRNA display.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6594)
  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 6594)
  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: plIVMD115
FEATURES             Location/Qualifiers
     source          1..6594
                     /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             1467..1496
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1502..1518)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1503..1519)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1521..1768
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1537..1555)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1786..1805)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1787..1805)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1815..1832)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1815..1831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1824..1846)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1972..2427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2059..2078)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2269..2290
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2558..3361)
                     /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(3362..3577)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3601..3620)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3720..3742
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3780..3798)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3839..4342
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4379..4483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4484..4552,4553..5344)
                     /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(4702..4721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5515..6103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6004..6023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6257..6274
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6391..6412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6427..6432,6433..6450,6451..6457)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6465..6481
                     /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     6470..6492
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6489..6505
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6489..6505
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6524..6544
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6526..6544
                     /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 cccttcgaac aaaaactaat ctcagaagag gatctgtaaa
     1501 tcgataccgt cgacctcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     1561 ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     1621 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     1681 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     1741 tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg
     1801 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1861 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1921 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     1981 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2041 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2101 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2161 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2221 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2281 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2341 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2401 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     2461 cggtatttca caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg
     2521 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     2581 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     2641 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     2701 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     2761 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     2821 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     2881 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     2941 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     3001 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     3061 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     3121 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     3181 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     3241 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     3301 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     3361 tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc
     3421 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     3481 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     3541 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag
     3601 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     3661 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3721 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     3781 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgga
     3841 tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt
     3901 tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag
     3961 gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa
     4021 aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag atatacattc
     4081 gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac
     4141 aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat
     4201 ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt
     4261 aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat
     4321 aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg
     4381 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4441 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4501 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4561 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4621 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4681 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4741 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4801 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4861 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4921 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4981 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5041 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5101 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5161 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5221 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5281 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5341 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5401 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5461 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5521 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5581 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5641 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5701 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5761 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5821 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5881 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5941 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6001 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6061 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6121 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6181 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6241 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6301 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6361 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6421 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6481 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     6541 aagggaacaa aagctggagc tccggatgca agggttcgaa tcccttagct ctca
//
