LOCUS       sequence-199238-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli/S. cerevisiae amdS shuttle vector allowing cloning of 
            ribozyme flanked g-RNA for Cas9 editing (HH-gRNA-HDV).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  AUTHORS   de Vries ARG, de Groot PA, van den Broek M, Daran JG
  TITLE     CRISPR-Cas9 mediated gene deletions in lager yeast Saccharomyces 
            pastorianus.
  JOURNAL   Microb Cell Fact. 2017 Dec 5;16(1):222. doi: 
            10.1186/s12934-017-0835-1.
  PUBMED    29207996
REFERENCE   2  (bases 1 to 5867)
  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: "Microb Cell
            Fact."; date: "2017-12-5"; volume: "16(1):222. doi"; pages: " 
            10.1186/s12934-017-0835-1"
COMMENT     Sequence Label: pUDP003
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2868..3534
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3404..3427
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(3596..3614)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(join(4048..4839,4840..4908))
                     /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     4671..4690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4909..5013)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5116..5704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5605..5624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
       61 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      121 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
      181 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      241 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      301 ttgctaggat acagttctca catcacatcc gaacataaac aaggatccat gccacaatct
      361 tgggaagaat tggctgctga caagagagct agattggcta agactatccc agacgaatgg
      421 aaggttcaaa ctttgccagc tgaagactct gttatcgact tcccaaagaa gtctggtatc
      481 ttgtctgaag ctgaattgaa gatcactgaa gcttctgctg ctgacttggt ttctaagttg
      541 gctgctggtg aattgacttc tgttgaagtt actttggctt tctgtaagag agctgctatc
      601 gctcaacaat tgactaactg tgctcacgaa ttcttcccag acgctgcttt ggctcaagct
      661 agagaattgg acgaatacta cgctaagcac aaaagacctg ttggtccatt gcacggtttg
      721 ccaatctctt tgaaggacca attgagagtt aagggttacg aaacttctat gggttacatc
      781 tcttggttga acaagtacga cgaaggtgac tctgttttga ctactatgtt gagaaaggct
      841 ggtgctgttt tctacgttaa gacttctgtt ccacaaactt tgatggtttg tgaaactgtt
      901 aacaacatca tcggtagaac tgttaaccca agaaacaaga actggtcttg tggtggttct
      961 tctggtggtg aaggtgctat cgttggtatc agaggtggtg ttatcggtgt tggtactgac
     1021 atcggtggtt ctatcagagt tccagctgct ttcaacttct tgtacggttt aagaccatct
     1081 cacggtagat tgccatacgc taagatggct aactctatgg aaggtcaaga aactgtacac
     1141 tctgttgttg gtccaatcac tcactctgtt gaagacttga gattgttcac taagtctgtt
     1201 ttgggtcaag aaccatggaa gtacgactct aaggttatcc caatgccatg gagacaatct
     1261 gaatctgaca tcatcgcttc taagatcaag aacggtggtt tgaacatcgg ttactacaac
     1321 ttcgacggta acgttttgcc acacccacca atcttgagag gtgttgaaac tactgttgct
     1381 gctttggcta aggctggtca cactgttact ccatggactc catacaagca cgacttcggt
     1441 cacgacttga tctctcacat ctacgctgct gacggttctg ctgacgttat gagagacatc
     1501 tctgcttctg gtgaaccagc tatcccaaac atcaaggact tgttgaaccc aaacatcaag
     1561 gctgttaaca tgaacgaatt gtgggacact cacttgcaaa agtggaacta ccaaatggaa
     1621 tacttggaaa agtggagaga agctgaagaa aaggctggta aggaattgga cgctatcatc
     1681 gctccaatca ctccaactgc tgctgttaga cacgaccaat tcagatacta cggttacgct
     1741 tctgttatca acttgttgga cttcacttct gttgttgttc cagttacttt cgctgacaag
     1801 aacatcgaca agaagaacga atctttcaag gctgtttctg aattggacgc tttggttcaa
     1861 gaagaatacg acccagaagc ttaccacggt gctccagttg ctgttcaagt tatcggtaga
     1921 agattgtctg aagaaagaac tttggctatc gctgaagaag ttggtaagtt gttgggtaac
     1981 gttgttactc cataactcga ggacaataaa aagattcttg ttttcaagaa cttgtcattt
     2041 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     2101 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     2161 gcgtcaatcg tatgtgaatg ctggtcgcta tactggagct ctcgagaacc ctcacctttc
     2221 gagaggacga tgcccgtgtc taaatgattc gaccagccta agaatgttca actcaacatc
     2281 tttggataat atcagaatga gaaagaacag atacgcagta cgttttttgg tgagctcttt
     2341 gcacttcttt agttctttcc atcaatatca gttttttaaa cttttaggac taaaagtgat
     2401 gtttaacttc aaaatgttta aaattttgtt cttcccgacg ttcattaaga atactaatac
     2461 actttaataa ttattttaat attttgttct aaataatgac ttttaattaa aaaagataaa
     2521 atataaaaac atcataataa ctcaccagag gttaagaaca aaaaaacaaa ttagatatct
     2581 gctaatccaa tatagttaaa tcaatctttc cttggtataa tgggtatatt acatatattt
     2641 caaggaccga cactcctacc aaatatctaa aatttaccat attaacataa catgtatata
     2701 aacgtcaaat cataatcagc actaacgtct cacggatcgt atatgccgta gcgacaatct
     2761 aagaactatg cgaggacacg ctagggccaa ggcaaaaaga ttccttgatt acgtaaggga
     2821 gttagaatca ttttgaataa aaaacacgct ttttcagttc gagtttatca ttatcaatac
     2881 tgccatttca aagaatacgt aaataattaa tagtagtgat tttcctaact ttatttagtc
     2941 aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta
     3001 cacagaatat ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa
     3061 tataatggag cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa
     3121 tattgttttc ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga
     3181 acaggggcac aaacaggcaa aaaacgggca caacctcaat ggagtgatgc aacctgcctg
     3241 gagtaaatga tgacacaagg caattgaccc acgcatgtat ctatctcatt ttcttacacc
     3301 ttctattacc ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt
     3361 tccctgaaat tattccccta cttgactaat aagtatataa agacggtagg tattgattgt
     3421 aattctgtaa atctatttct taaacttctt aaattctact tttatagtta gtcttttttt
     3481 tagttttaaa acaccaagaa cttagtttcg aataaacaca cataaacaaa caaacgagac
     3541 ctgcggaggt ctccacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac
     3601 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     3661 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     3721 tatttcaaat ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac attatactga
     3781 aaaccttgct tgagaaggtt ttgggacaag cttacgtctc acggatcgta tatgccgtag
     3841 cgacaatcta agaactatgc gaggacacgc tagcagctgt attcacgtag acggataggt
     3901 atagccagac atcagcagca tacttcggga accgtaggcc accgagcggc cgcgattatc
     3961 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4021 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4081 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4141 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
     4201 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4261 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4321 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4381 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4441 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4501 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4561 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4621 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4681 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4741 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4801 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4861 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4921 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4981 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgt
     5041 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5221 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5761 catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggact atatgtgaag
     5821 gcatggctat ggcacggcag acattccgcc agatcatcaa taggcac
//
