LOCUS       sequence-478644-        5197 bp    DNA     circular SYN 15-JUL-2025
DEFINITION  Nb Library Receiver Vector (under galactose induction), AmpR (E. 
            coli) and TRP1 (yeast), Cen6/ARS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5197)
  TITLE     High-throughput Activity Reprogramming of Protease (HARP)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pY3-NbLibRec
FEATURES             Location/Qualifiers
     source          1..5197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1276..1295)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1277..1295)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1306..1970
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(1319..1342)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1637..1754
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     rep_origin      complement(2383..2971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2463..2482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3074..3178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3179..3247,3248..4039)
                     /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(3397..3416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_feature    4150..4653
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4694..4712
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4750..4772)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4872..4891
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4908..5188
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             join(5189..5197,1..666)
                     /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"
ORIGIN
        1 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
       61 gccgcagaat gtgctctaga ttctgatgct gacttgctgg gtattatatg tgtgcccaat
      121 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
      181 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
      241 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
      301 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
      361 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
      421 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
      481 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
      541 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
      601 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
      661 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg cctgcggtgt
      721 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggtgtactgc agtgcactag
      781 tgaattggag ctcaattctc ttaggattcg attcacattc atcttttttt agctactaat
      841 aggataaatt ataggaattt ataacaaatt gaaatatggc aggcagcaaa attaaaattg
      901 tcttagtttt ttagtgtata gaagtgaata gctatataaa gtatgtgtaa agttggtaac
      961 ggaacgaaaa atagaaaagg atattacatg ggaaaacatg ttgtttacgg agaaatgaaa
     1021 agtatattgt attttgtacg agctaaaagt acagtgggaa caaagtcgat tttgttacat
     1081 ctacactgtt gttatcagat ctcgagctat tactagccgc ggtaccaagc ttaatgcatg
     1141 cttagtctta agctgccgtc gacagctgcc gcccgcctgc accaggccgc cgccgctttc
     1201 ctgcagctgc acctgtgcta aaactgaagc aataacagaa aatattgaaa aacagcgaag
     1261 taactgcatg ctagccccta tagtgagtcg tattacggat ccggggtttt ttctccttga
     1321 cgttaaagta tagaggtata ttaacaattt tttgttgata cttttattac atttgaataa
     1381 gaagtaatac aaaccgaaaa tgttgaaagt attagttaaa gtggttatgc agtttttgca
     1441 tttatatatc tgttaataga tcaaaaatca tcgcttcgct gattaattac cccagaaata
     1501 aggctaaaaa actaatcgca ttatcatcct atggttgtta atttgattcg ttcatttgaa
     1561 ggtttgtggg gccaggttac tgccaatttt tcctcttcat aaccataaaa gctagtattg
     1621 tagaatcttt attgttcgga gcagtgcggc gcgaggcaca tctgcgtttc aggaacgcga
     1681 ccggtgaaga cgaggacgca cggaggagag tcttccttcg gagggctgtc acccgctcgg
     1741 cggcttctaa tccgtacttc aatatagcaa tgagcagtta agcgtattac tgaaagttcc
     1801 aaagagaagg tttttttagg ctaagataat ggggctcttt acatttccac aacatataag
     1861 taagattaga tatggatatg tatatggata tgtatatggt ggtaatgcca tgtaatatga
     1921 ttattaaact tctttgcgtc catccaaaaa aaaagtaaga atttttgaaa attcgaattc
     1981 aaccctcact aaagtctgac taatatcatc ggtataagtc ggtgccgcgt accatggtct
     2041 atacatgggt gaccaaaaga gcgggcggat acacgcgtca ccacaagcag aataaaaggt
     2101 aaacctgaaa ttgttttaac ataaaatgaa aaatgcttgt ttgcaaccct atatagaatc
     2161 ataaaacatt cgtgactata aaatgaataa actaaactat tctaagaaaa tgaaataaat
     2221 gacaaaaaaa cgtgtttttt ggactagaag gcttaatcaa aagctgaatt caggggcggc
     2281 cgcacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2821 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3001 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgacag gtggcacttt
     3061 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3121 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3181 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3241 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3301 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3361 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3421 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3481 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3541 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3601 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3661 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3721 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3781 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3841 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtcccg
     3901 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3961 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4021 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4081 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atcgcggccg
     4141 ctcgggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
     4201 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
     4261 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
     4321 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
     4381 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggttt
     4441 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
     4501 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
     4561 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
     4621 atttaaatta taattatttt tatagcacgt gatactctaa gaaaccatta ttatcatgac
     4681 attaacctat aaaaataggc gtatcacgag gccctttcga ctcgcgcgtt tcggtgatga
     4741 cggtgaaaac ctctgacaca tgcagctccc ggacacggtc acagcttgtc tgtaagcgga
     4801 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4861 gcttaactat gcggcatcag agcagattgt actgagagtg caccataaac gacattacta
     4921 tatatataat ataggaagca tttaatagac agcatcgtaa tatatgtgta ctttgcagtt
     4981 atgacgccag atggcagtag tggaagatat tctttattga aaaatagctt gtcaccttac
     5041 gtacaatctt gatccggagc ttttcttttt ttgccgatta agaattaatt cggtcgaaaa
     5101 aagaaaagga gagggccaag agggagggca ttggtgacta ttgagcacgt gagtatacgt
     5161 gattaagcac acaaaggcag cttggagtat gtctgtt
//
