LOCUS       sequence-136121-        5808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains the CORE cassette with KlURA3 and hyg markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5808)
  AUTHORS   Storici F, Resnick MA
  TITLE     Delitto perfetto targeted mutagenesis in yeast with 
            oligonucleotides.
  JOURNAL   Genet Eng (N Y). 2003;25:189-207.
  PUBMED    15260239
REFERENCE   2  (bases 1 to 5808)
  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: "Genet Eng 
            (N Y)."; date: "2003"; volume: "25"; pages: "189-207"
COMMENT     Sequence Label: pCORE-UH
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        237..255
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            complement(1843..3418)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(1843..2040)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2046..3074)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(2046..3074)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3075..3418)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(3551..3568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3722..4310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3802..3821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4481..5272,5273..5341))
                     /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     5104..5123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5342..5446)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5514..5532
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5570..5592)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5692..5711
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5792..5808,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5792..5808,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt
      241 aaccctcact aaagggaaca aaagctggta ccgggcccgg agacaatcat atgggagaag
      301 caattggaag atagaaaaaa ggtactcggt acataaatat atgtgattct gggtagaaga
      361 tcggtctgca ttggatggtg gtaacgcatt tttttacaca cattacttgc ctcgagcatc
      421 aaatggtggt tattcgtgga tctatatcac gtgatttgct taagaattgt cgttcatggt
      481 gacactttta gctttgacat gattaagctc atctcaattg atgttatcta aagtcatttc
      541 aactatctaa gatgtggttg tgattgggcc attttgtgaa agccagtacg ccagcgtcaa
      601 tacactcccg tcaattagtt gcaccatgtc cacaaaatca tataccagta gagctgagac
      661 tcatgcaagt ccggttgcat cgaaactttt acgtttaatg gatgaaaaga agaccaattt
      721 gtgtgcttct cttgacgttc gttcgactga tgagctattg aaactagttg aaacgttggg
      781 tccatacatt tgccttttga aaacacacgt tgatatcttg gatgatttca gttatgaggg
      841 tactgtcgtt ccattgaaag cattggcaga gaaatacaag ttcttgatat ttgaggacag
      901 aaaattcgcc gatatcggta acacagtcaa attacaatat acatcgggcg tttaccgtat
      961 cgcagaatgg tctgatatca ccaacgccca cggggttact ggtgctggta ttgttgctgg
     1021 cttgaaacaa ggtgcgcaag aggtcaccaa agaaccaagg ggattattga tgcttgctga
     1081 attgtcttcc aagggttctc tagcacacgg tgaatatact aagggtaccg ttgatattgc
     1141 aaagagtgat aaagatttcg ttattgggtt cattgctcag aacgatatgg gaggaagaga
     1201 agaagggttt gattggctaa tcatgacccc aggtgtaggt ttagacgaca aaggcgatgc
     1261 attgggtcag cagtacagaa ccgtcgacga agttgtaagt ggtggatcag atatcatcat
     1321 tgttggcaga ggacttttcg ccaagggtag agatcctaag gttgaaggtg aaagatacag
     1381 aaatgctgga tgggaagcgt accaaaagag aatcagcgct ccccattaat tatacaggaa
     1441 acttaataga acaaatcaca tatttaatct aatagccacc tgcattggca cggtgcaaca
     1501 ctcacttcaa cttcatctta caaaagatca cgtgatctgt tgtattgaac tgaaaatttt
     1561 ttgtttgctt ctctctctct ctttcattat gtgagagttt aaaaaccaga aactacatca
     1621 tcgaaaaaga gtttaaacca ttacaaccat tgcgataagc cctctcaaac ttcctccaga
     1681 ctgtttgtca tccaattggt aagattaatt atcatatacc ccggataaga aaagaagcaa
     1741 gcacacaatg gatcaactta atggtaagga acaacaagcg gccgccaccg cggtggagct
     1801 cgttttcgac actggatggc ggcgttagta tcgaatcgac agcagtatag cgaccagcat
     1861 tcacatacga ttgacgcatg atattacttt ctgcgcactt aacttcgcat ctgggcagat
     1921 gatgtcgagg cgaaaaaaaa tataaatcac gctaacattt gattaaaata gaacaactac
     1981 aatataaaaa aactatacaa atgacaagtt cttgaaaaca agaatctttt tattgtcagt
     2041 actgattatt cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt
     2101 acttctacac agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg
     2161 acagtcccgg ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca
     2221 tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac
     2281 gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc
     2341 tgctccatac aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa
     2401 tccccgaaca tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg
     2461 acattgttgg agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc
     2521 caaagcatca gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca
     2581 gtttgccagt gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg
     2641 tattgaccga ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc
     2701 gcagcgatcg catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca
     2761 ggcaggtctt gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg
     2821 ctgaattccc caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga
     2881 taaacataac gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca
     2941 cgccctccta catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg
     3001 tcggagacgc tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca
     3061 ggctttttac ccatggttgt ttatgttcgg atgtgatgtg agaactgtat cctagcaaga
     3121 ttttaaaagg aagtatatga aagaagaacc tcagtggcaa atcctaacct tttatatttc
     3181 tctacagggg cgcggcgtgg ggacaattca acgcgtctgt gaggggagcg tttccctgct
     3241 cgcaggtccg cagcgaggag ccgtaatttt tgcttcgcgc cgtgcggcca tcaaaatgta
     3301 tggatgcaaa tgattataca tggggatgta tgggctaaat gtacgggcga cagtcacatc
     3361 atgcccctga gctgcgcacg tcaagactgt caaggagggt attctgggcc tccatgtcgc
     3421 tggccgggtg acccggcggg gacgaggcaa gctaaacaga tctggcgcgc cttaattaac
     3481 ccaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3901 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4141 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5461 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5521 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5581 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     5641 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     5701 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     5761 aatacataac cttatgtatc atacacatac gatttaggtg acactata
//
