LOCUS       sequence-317191-        6708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo mRNA display construct for mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6708)
  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 6708)
  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: plIVMD163
FEATURES             Location/Qualifiers
     source          1..6708
                     /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"
     protein_bind    744..768
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             774..1481
                     /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(919..937)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1187..1206
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1483..1507)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1527..1544
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1635..1882
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1651..1669)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1900..1919)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1901..1919)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1929..1946)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1929..1945)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1938..1960)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2086..2541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2173..2192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2383..2404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2672..3475)
                     /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(3476..3691)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3715..3734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3834..3856
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3894..3912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3953..4456
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4493..4597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4598..4666,4667..5458)
                     /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(4816..4835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5629..6217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6118..6137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6371..6388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6505..6526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6541..6546,6547..6564,6565..6571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6579..6595
                     /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     6584..6606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6603..6619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6603..6619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6638..6658
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6640..6658
                     /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 accacaagtt tgtacaaaaa agcaggctac aaaatggtga
      781 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      841 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      901 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      961 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     1021 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1081 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1141 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1201 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1261 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1321 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1381 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1441 agggccgcca ctccaccggc ggcatggacg agctgtacaa gaacccagct ttcttgtaca
     1501 aagtggtttc aagagggccc ttcgaacatc atcaccatca ccattaaatc gataccgtcg
     1561 acgatcctac ggtacttatt gccaagaaag cacgagcatc agccgtgcct ccaggtcgaa
     1621 tcttcaaact cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac
     1681 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     1741 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     1801 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     1861 acgctcgaag gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac
     1921 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1981 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2041 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     2101 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2161 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2221 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2281 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2341 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2401 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2461 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2521 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2581 ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt tgtgagttta
     2641 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     2701 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     2761 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     2821 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     2881 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     2941 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     3001 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     3061 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     3121 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     3181 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     3241 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     3301 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     3361 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     3421 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     3481 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     3541 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     3601 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa
     3661 aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat
     3721 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3781 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3841 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3901 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cggatcgctt
     3961 gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct
     4021 gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa
     4081 gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt
     4141 actttacata tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa
     4201 cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg
     4261 aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg
     4321 gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc
     4381 tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat
     4441 ttttatagca cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac
     4501 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4561 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4621 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4681 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4741 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4801 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4861 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4921 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4981 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5041 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5101 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5161 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5221 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5281 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5341 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5401 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5461 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5521 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5761 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6601 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     6661 acaaaagctg gagctccgga tgcaagggtt cgaatccctt agctctca
//
