LOCUS       sequence-339123-        4638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P1 integration cassette for AT110 Polymerase for integration onto 
            Orthorep for Nanobody evolution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4638)
  AUTHORS   Wellner A, McMahon C, Gilman MSA, Clements JR, Clark S, Nguyen KM, 
            Ho MH, Hu VJ, Shin JE, Feldman J, Hauser BM, Caradonna TM, Wingler 
            LM, Schmidt AG, Marks DS, Abraham J, Kruse AC, Liu CC
  TITLE     Rapid generation of potent antibodies by autonomous hypermutation in
            yeast.
  JOURNAL   Nat Chem Biol. 2021 Jun 24. pii: 10.1038/s41589-021-00832-4. doi: 
            10.1038/s41589-021-00832-4.
  PUBMED    34168368
REFERENCE   2  (bases 1 to 4638)
  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 Chem 
            Biol. 2021 Jun 24. pii: 10.1038/s41589-021-00832-4. doi: 
            10.1038/s41589-021-00832-4."
COMMENT     Sequence Label: pAW240
FEATURES             Location/Qualifiers
     source          1..4638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     371..388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1335..1346
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1347..1373
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_RNA        1744..1795
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      1802..1967
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1857..1881)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2039..2710)
                     /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"
     primer_bind     2999..3021
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        complement(3961..4052)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4119..4166)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4162..4180
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     rep_origin      join(4267..4638,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gagcgcaacg caatagtact ataatatatg aattacatta ttaatttaaa
      541 aacgcgtgat gacctataca taggaagatc tatagaaaca aaaagattaa taactttcaa
      601 atatcagaaa aatatagaaa catgtgataa gctcatagac atataaagga tccatgagat
      661 tcccatctat cttcaccgct gctttgttcg ctgcttcttc tgctttggct gctccagcta
      721 acaccaccac cgaagacgaa accgctcaaa tcccagctga agctgttatc gactactctg
      781 acttggaagg tgacttcgac gctgctgctt tgccattgtc taactctacc aacaacggtt
      841 tgtcttctac caacaccacc atcgcttcta tcgctgctaa ggaagaaggt gttcaattgg
      901 acaagagaga agctgacgca caggtgcact cctcgctgca ggaaagcggc ggcggcctgg
      961 tgcaggcggg cggcagcctg cgcctgagct gcgcggcgag cggcaatatt tttgacgctg
     1021 acatcatggg ctggtatcgc caggcgccgg gcaaagaacg cgaacttgtt gccagtatta
     1081 ctgacggtgg tagtaccaat tatgcggata gcgtgaaagg ccgctttacc attagccgcg
     1141 ataacgcgaa aaacaccgtg tatctgcaga tgaacagcct gaaaccggaa gataccgcgg
     1201 tgtattattg cgcggctatc gcttacccgg acatcccgac ttacttcgac tacgactctg
     1261 actatttcta ttggggccag ggcacccagg tgaccgtgag aggaggcagc ggatccaagg
     1321 acaatagctc gacgattgaa ggtagatacc catacgacgt tccagactac gctctgcagg
     1381 ctagtggtgg tggtggttct ggtggtggtg gttctggtgg tggtggttct gctagcgacg
     1441 tccaggaact gacaactata tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt
     1501 actctttgtc aacgactact attttggcca acgggaaggc aatgcaagga gtttttgaat
     1561 attacaaatc agtaacgttt gtcagtaatt gcggttctca cccctcaaca actagcaaag
     1621 gcagccccat aaacacacag tatgtttttt gaagaagact tgtcatgaaa aaaaaaaaaa
     1681 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1741 aaacctgtca ccggatgtgt tttccggtct gatgagtccg tgaggacgaa acagggaatt
     1801 cgcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
     1861 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
     1921 tttcctgtag gtcaggttgc tttctcaggt atagcatgag gtcgctcgag ctccgtttct
     1981 attatgaatt tcatttataa agtttatgta caaatatcat aaaaaaagag aatctttttt
     2041 tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca
     2101 cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt
     2161 ctggcgtcag tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa
     2221 cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa
     2281 acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt
     2341 ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg
     2401 actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat
     2461 cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt
     2521 tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc
     2581 tattgggcac acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg
     2641 cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa
     2701 taacagacat tttataatta ttataaaaac tttcatataa gagatataaa atttaatatg
     2761 gaagaaataa gacaagaaag atacaaccaa atgaaagaag ctctaaatag tgttgaaggt
     2821 tataaaggaa aaattgtagc ctcagactca gattggtgtt tcaaagatcc tcaaggcaat
     2881 agaataacag attttgatag tactttaagc cagccccgac acccgccaac acccgctgac
     2941 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3001 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaaggca
     3061 aaaaaaaagg ctccaaaagg agcctttaat tgtatcggtt taccaatgct taatcagtga
     3121 agcccctatt tcagcaatct gtctatttcg ttcgtccata gttgcctgac tacccgtcgt
     3181 gtagataact acgatacggg aaggcttacc atctggtcca agtgcggcaa tgataccccg
     3241 tgaaccacgc tctcctgccc cagatttatc agcaataaac cagccagcag gaagtgccga
     3301 acgcagtagt ggtcctgcaa ctttatccgc ttcaagccag tctattaatt gttgccgtga
     3361 agccagagta agtagttcgc cagttaatag tttgcgtaat gttgttgcca ttgccgcagg
     3421 cattgtggtg tctcgctcgt cgtttggtat ggcttcattc agttctggtt cccaacggtc
     3481 aaggcgagtt acatggtctc ccatattgtg caaaaaagcg gttaattcct tcggtccacc
     3541 gatggttgtc agaagtaagt tcgccgcagt gttatcactc atcgttatgg cggcactaca
     3601 taattctctt accgtcattc cgtccgtaag gtgcttttct gtcactgggg agtattcaac
     3661 caagtcattc tgcgaatagt gtattcgtcg tccgagttgc tcttgccccg catctacacg
     3721 ggataatact gctccacata gcagaacttt gaaagtgctc atcattggaa aacgctcttc
     3781 agggcgaaaa ctctcaagga ttttaccgct attcaagtcc agttctatgt atcccactcg
     3841 tgcccccaat tggtcctctg cgtcttttac tttcacgagc gtttctgggt gagcaaaaac
     3901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctga tgagcggata
     4021 catatttgaa tgtatttaga aaaataaaca aataggggaa ttaaaaaaaa gcccgctcat
     4081 taggcgggct actagtcgga tatagttcct cctttcagca aaaaacccct caagacccgt
     4141 ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct
     4201 tcctttcggg ctttgttagc agccggatct tctagaagac cccgtagaaa agatcaaagg
     4261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4381 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4621 aacgacctac accgaact
//
