LOCUS       sequence-136067-        5589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains the CORE cassette with convergent KlURA3 and KanMX4 
            markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5589)
  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 5589)
  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-UK
FEATURES             Location/Qualifiers
     source          1..5589
                     /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..3199)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(1843..2040)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2046..2855)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2769..2788
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2856..3199)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(3332..3349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3503..4091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3583..3602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4262..5053,5054..5122))
                     /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     4885..4904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5123..5227)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5295..5313
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5351..5373)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5473..5492
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5573..5589,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5573..5589,1)
                     /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 ctacttcaac ttcatcttac aaaaagatca cgtgatctgt tgtattgaac tgaaaatttt
     1561 ttgtttgctt ctctctctct ctttcattat gtgagattta aaaaccagaa actacatcat
     1621 cgaaaaagag ttttaaacca 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 actgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     2101 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2161 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2221 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2281 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     2341 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     2401 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     2461 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     2521 ggatattctt ctaatacctg gaatgctgtt ttgccgggga tcgcagtggt gagtaaccat
     2581 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     2641 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2701 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2761 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2821 cgcggcctcg aaacgtgagt cttttcctta cccatggttg tttatgttcg gatgtgatgt
     2881 gagaactgta tcctagcaag attttaaaag gaagtatatg aaagaagaac ctcagtggca
     2941 aatcctaacc ttttatattt ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg
     3001 tgaggggagc gtttccctgc tcgcaggtct gcagcgagga gccgtaattt ttgcttcgcg
     3061 ccgtgcggcc atcaaaatgt atggatgcaa atgattatac atggggatgt atgggctaaa
     3121 tgtacgggcg acagtcacat catgcccctg agctgcgcac gtcaagactg tcaaggaggg
     3181 tattctgggc ctccatgtcg ctggccgggt gacccggcgg ggacgaggca agctaaacag
     3241 atctggcgcg ccttaattaa cccaacctgc attaatgaat cggccaacgc gcggggagag
     3301 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3361 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3421 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3481 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3541 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3601 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3661 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     3721 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3781 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3841 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3901 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3961 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4021 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4081 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4141 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4201 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4261 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4321 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4381 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4441 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4501 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4561 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4621 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4681 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4741 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4801 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4861 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4921 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4981 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5041 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5101 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5161 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5221 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5281 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5341 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5401 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     5461 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt
     5521 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cgatttaggt
     5581 gacactata
//
