LOCUS       sequence-338565-        4614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P1 integration cassette for AT110 Polymerase for integration onto 
            Orthorep for Nanobody evolution with different restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4614)
  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 4614)
  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: pAW212
FEATURES             Location/Qualifiers
     source          1..4614
                     /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             1311..1322
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1323..1349
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_RNA        1720..1771
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      1778..1943
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1833..1857)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2015..2686)
                     /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     2975..2997
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        complement(3937..4028)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4095..4142)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4138..4156
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     rep_origin      join(4243..4614,1..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 gttttgttcg 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 ggtacccagg tgcagctgca ggaaagcggc ggcggcctgg
      961 tgcaggcggg cggcagcctg cgcctgagct gcgcggcgag cggctatatt tctgacgctt
     1021 actacatggg ctggtatcgc caggcgccgg gcaaagaacg cgaatttgtt gccactatta
     1081 ctcatggtac taatacctat tatgcggata gcgtgaaagg ccgctttacc attagccgcg
     1141 ataacgcgaa aaacaccgtg tatctgcaga tgaacagcct gaaaccggaa gataccgcgg
     1201 tgtattattg cgcggttctg gaaactcgtt cttactcttt tcgttattgg ggccagggca
     1261 cccaggtgac cgtgagcagc gtcgacggtt ccaaggacaa tagctcgacg attgaaggta
     1321 gatacccata cgacgttcca gactacgctc tgcaggctag tggtggtggt ggttctggtg
     1381 gtggtggttc tggtggtggt ggttctgcta gcgacgtcca ggaactgaca actatatgcg
     1441 agcaaatccc ctcaccaact ttagaatcga cgccgtactc tttgtcaacg actactattt
     1501 tggccaacgg gaaggcaatg caaggagttt ttgaatatta caaatcagta acgtttgtca
     1561 gtaattgcgg ttctcacccc tcaacaacta gcaaaggcag ccccataaac acacagtatg
     1621 ttttttgaag aagacttgtc atgaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1681 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaac ctgtcaccgg atgtgttttc
     1741 cggtctgatg agtccgtgag gacgaaacag ggaattcgcg aatttcttat gatttatgat
     1801 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     1861 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     1921 tcaggtatag catgaggtcg ctcgagctcc gtttctatta tgaatttcat ttataaagtt
     1981 tatgtacaaa tatcataaaa aaagagaatc tttttttctt agcatttttg acgaaatttg
     2041 ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa
     2101 taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca
     2161 taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat
     2221 ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt
     2281 tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa
     2341 ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc agttggacga
     2401 tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga ttacgaaaca
     2461 cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca agacttgaaa
     2521 ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat ataataccca
     2581 gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc aagccgcaaa
     2641 ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatttta taattattat
     2701 aaaaactttc atataagaga tataaaattt aatatggaag aaataagaca agaaagatac
     2761 aaccaaatga aagaagctct aaatagtgtt gaaggttata aaggaaaaat tgtagcctca
     2821 gactcagatt ggtgtttcaa agatcctcaa ggcaatagaa taacagattt tgatagtact
     2881 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2941 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3001 tcaccgtcat caccgaaacg cgcgagacga aaggcaaaaa aaaaggctcc aaaaggagcc
     3061 tttaattgta tcggtttacc aatgcttaat cagtgaagcc cctatttcag caatctgtct
     3121 atttcgttcg tccatagttg cctgactacc cgtcgtgtag ataactacga tacgggaagg
     3181 cttaccatct ggtccaagtg cggcaatgat accccgtgaa ccacgctctc ctgccccaga
     3241 tttatcagca ataaaccagc cagcaggaag tgccgaacgc agtagtggtc ctgcaacttt
     3301 atccgcttca agccagtcta ttaattgttg ccgtgaagcc agagtaagta gttcgccagt
     3361 taatagtttg cgtaatgttg ttgccattgc cgcaggcatt gtggtgtctc gctcgtcgtt
     3421 tggtatggct tcattcagtt ctggttccca acggtcaagg cgagttacat ggtctcccat
     3481 attgtgcaaa aaagcggtta attccttcgg tccaccgatg gttgtcagaa gtaagttcgc
     3541 cgcagtgtta tcactcatcg ttatggcggc actacataat tctcttaccg tcattccgtc
     3601 cgtaaggtgc ttttctgtca ctggggagta ttcaaccaag tcattctgcg aatagtgtat
     3661 tcgtcgtccg agttgctctt gccccgcatc tacacgggat aatactgctc cacatagcag
     3721 aactttgaaa gtgctcatca ttggaaaacg ctcttcaggg cgaaaactct caaggatttt
     3781 accgctattc aagtccagtt ctatgtatcc cactcgtgcc cccaattggt cctctgcgtc
     3841 ttttactttc acgagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3901 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3961 aagcatttat cagggttatt gtctgatgag cggatacata tttgaatgta tttagaaaaa
     4021 taaacaaata ggggaattaa aaaaaagccc gctcattagg cgggctacta gtcggatata
     4081 gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     4141 agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt gttagcagcc
     4201 ggatcttcta gaagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4261 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4321 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4381 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4441 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4501 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4561 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aact
//
