LOCUS       sequence-430590-        7201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast surface display Fab vector with lambda light chain and mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7201)
  AUTHORS   Petersen BM, Kirby MB, Chrispens KM, Irvin OM, Strawn IK, Haas CM, 
            Walker AM, Baumer ZT, Ulmer SA, Ayala E, Rhodes ER, Guthmiller JJ, 
            Steiner PJ, Whitehead TA
  TITLE     An integrated technology for quantitative wide mutational scanning 
            of human antibody Fab libraries.
  JOURNAL   Nat Commun. 2024 May 10;15(1):3974. doi: 10.1038/s41467-024-48072-z.
  PUBMED    38730230
REFERENCE   2  (bases 1 to 7201)
  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: "Nat 
            Commun."; date: "2024-05-10"; volume: "15(1):3974. doi"; pages: " 
            10.1038/s41467-024-48072-z"
COMMENT     Sequence Label: pYSD_lambda_mRFP
FEATURES             Location/Qualifiers
     source          1..7201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        397..501
                     /gene="bla"
                     /label=AmpR promoter
     CDS             502..1317
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             502..1317
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(575..594)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1496..2084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1985..2004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2238..2255
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2372..2393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2408..2413,2414..2431,2432..2438)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2446..2462
                     /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     2451..2473
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2470..2486
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2470..2486
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2507..2525
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(2567..2732)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2653..2677
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2886..2927)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3825..3854
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4183..4202)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4184..4202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4209..4226)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4209..4225)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4218..4240)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    4366..4384
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4391..4409
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4427..4438
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4445..5122
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      5140..5211
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5227..5254
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      5286..5741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5373..5392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5583..5604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5844..6518)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(6519..6799)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6820..6839)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6939..6961
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6999..7017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(7058..7201,1..360)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
       61 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
      121 aaatcacagg attttcgtgt gtggtcgtct acacagacaa gatgaaacaa ttcggcatta
      181 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
      241 cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat
      301 ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat
      361 gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      421 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      481 taatattgaa aaaggaagag tatgagccat attcaacggg aaacgtcttg ctctaggccg
      541 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
      601 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
      661 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
      721 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
      781 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
      841 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
      901 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
      961 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1021 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     1081 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1141 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1201 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1261 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaacct
     1321 cagcaactgt caaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1381 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     2281 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     2341 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     2401 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     2461 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt aaccctcact
     2521 aaagggaaca aaagctggta ccttcgctat tacgccagct gaattggagc gacctcatgc
     2581 tatacctgag aaagcaacct gacctacagg aaagagttac tcaagaataa gaattttcgt
     2641 tttaaaacct aagagtcact ttaaaatttg tatacactta ttttttttat aacttattta
     2701 ataataaaaa tcataaatca taagaaattc gcttatttag aagtgtcaac aacgtatcta
     2761 ccaacgattt gacccttttc catcttttcg taaatttctg gcaaggtaga caagccgaca
     2821 accttgattg gagacttgac caaacctctg gcgaagaatt gttaattaag agctcagatc
     2881 tattaggtgg aatccaggcc cagcagcggg ttcggaatcg gtttgccgct gcactcggtg
     2941 ggggccacgg tcttttccac ggtgctgccc tcgtgggtca cctggcagct gtagctccgg
     3001 tggctcttcc actgctcggg ggtgaggctc aggtagctgc tggcggcgta cttgttgttg
     3061 ctctgcttgc tgggggtggt agtctccacg ccggccttca cggggctgct gtcggccttc
     3121 caggccacgg tcacggcgcc agggtagaag tcgctgatga ggcacaccag ggtggccttg
     3181 ttggcctgca gcggccgcag ctacacttca ttggatggat gaatcgcggc cgcccgcagg
     3241 tgttccccct ggcccccagc agcaagagca ccagcggcgg cacagccgcc ctgggctgcc
     3301 tggtgaagga ctacttcccc gagcccgtga ccgtgagctg gaacagcggc gccttgacca
     3361 gcggcgtgca caccttcccc gccgtgctgc agagcagcgg cctgtacagc ctgagcagcg
     3421 tggtgaccgt gcccagcagc agcctgggca cccagaccta catctgcaac gtgaaccaca
     3481 agcccagcaa caccaaggtg gacaaacgcg tggagcccaa gagctgcgat aagactcata
     3541 ccggtggtgg tggttctggt ggtggtggtt ctggtggtgg tggttctggt ggtggtggtt
     3601 ctcaggaact gacaactata tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt
     3661 actctttgtc aacgactact attttggcca acgggaaggc aatgcaagga gtttttgaat
     3721 attacaaatc agtaacgttt gtcagtaatt gcggttctca cccctcaaca actagcaaag
     3781 gcagccccat aaacacacag tatgtttttg gtgggggcgg atccgaacaa aagcttattt
     3841 ctgaagagga cttgtaatag agatctgata acaacagtgt agatgtaaca aaatcgactt
     3901 tgttcccact gtacttttag ctcgtacaaa atacaatata cttttcattt ctccgtaaac
     3961 aacatgtttt cccatgtaat atccttttct atttttcgtt ccgttaccaa ctttacacat
     4021 actttatata gctattcact tctatacact aaaaaactaa gacaatttta attttgctgc
     4081 ctgccatatt tcaatttgtt ataaattcct ataatttatc ctattagtag ctaaaaaaag
     4141 atgaatgtga atcgaatcct aagagaattg agctccaatt cgccctatag tgagtcgtat
     4201 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4261 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4321 accgatcgcc tttcccaaca gttgcgcagc cttgcctgag accagtccct atcagtgata
     4381 gagattgaca tccctatcag tgatagagat actgagcacg gatctgaaag aggagaaagg
     4441 atctatggcg agtagcgaag acgttatcaa agagttcatg cgtttcaaag ttcgtatgga
     4501 aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga
     4561 aggtactcag accgctaaac tgaaagttac caaaggtggt ccgctgccgt tcgcttggga
     4621 catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc cggctgacat
     4681 cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt
     4741 cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg gtgagttcat
     4801 ctacaaagtt aaactgcgtg gtactaactt cccgtccgac ggtccggtta tgcagaaaaa
     4861 aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg
     4921 tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg aagttaaaac
     4981 cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg acatcaaact
     5041 ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac gtgctgaagg
     5101 tcgtcactcc accggtgctt aataaggatc tccaggcatc aaataaaacg aaaggctcag
     5161 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     5221 cacactggct caccttcggg tgggcctttc tgcgtttata agtcggtctc aactatggcg
     5281 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5341 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5581 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5641 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5701 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     5761 cttacgcatc tgtgcggtat ttcacaccgc atagatcggc aagtgcacaa acaatactta
     5821 aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg
     5881 ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca
     5941 tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt
     6001 aagctttcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca atccaaaagt
     6061 tcacctgtcc ctcctgcttc tgaatcaaac aagggaataa acgaatgagg tttctgtgaa
     6121 gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat aactggcaaa
     6181 ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac gatatcaatg
     6241 ccgtaatcat tgaccagagc caaaacatcc tccttaggtt gattacgaaa cacgccaacc
     6301 aagtatttcg gagtgcctga actattttta tatgctttta caagacttga aattttcctt
     6361 gcaataaccg ggtcaattgt tctctttcta ttgggcacac atataatacc cagcaagtca
     6421 gcatcggaat ctagagcaca ttctgcggcc tctgtgctct gcaagccgca aactttcacc
     6481 aatggaccag aactacctgt gaaattaata acagacatac tccaagctgc ctttgtgtgc
     6541 ttaatcacgt atactcacgt gctcaatagt caccaatgcc ctccctcttg gccctctcct
     6601 tttctttttt cgaccgaatt aattcttaat cggcaaaaaa agaaaagctc cggatcaaga
     6661 ttgtacgtaa ggtgacaagc tatttttcaa taaagaatat cttccactac tgccatctgg
     6721 cgtcataact gcaaagtaca catatattac gatgctgttc tattaaatgc ttcctatatt
     6781 atatatatag taatgtcgtg atctatggtg cactctcagt acaatctgct ctgatgccgc
     6841 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6901 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6961 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     7021 ataggttaat gtcatgataa taatggtttc ttagacggat cgcttgcctg taacttacac
     7081 gcgcctcgta tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt
     7141 tatgttttgt atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa
     7201 g
//
