LOCUS       sequence-131660-        5299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains the CORE cassette with KlURA3  and KanMX4 markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5299)
  AUTHORS   Storici F, Lewis LK, Resnick MA
  TITLE     In vivo site-directed mutagenesis using oligonucleotides.
  JOURNAL   Nat Biotechnol. 2001 Aug;19(8):773-6.
  PUBMED    11479573
REFERENCE   2  (bases 1 to 5299)
  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 
            Biotechnol."; date: "2001-08"; volume: "19(8)"; pages: "773-6"
COMMENT     Sequence Label: pCORE
FEATURES             Location/Qualifiers
     source          1..5299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    59..80
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(95..100,101..118,119..125)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    133..149
                     /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     138..160
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     157..173
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     157..173
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        194..212
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1819..1837)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1924..1941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2095..2683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2175..2194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2854..3645,3646..3714))
                     /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     3477..3496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        3750..3768
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(3876..5232)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(3876..4073)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(4079..4888)
                     /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     4802..4821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4889..5232)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 gatccccctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
       61 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
      121 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
      181 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggtaccg ggcccggaga
      241 caatcatatg ggagaagcaa ttggaagata gaaaaaaggt actcggtaca taaatatatg
      301 tgattctggg tagaagatcg gtctgcattg gatggtggta acgcattttt ttacacacat
      361 tacttgcctc gagcatcaaa tggtggttat tcgtggatct atatcacgtg atttgcttaa
      421 gaattgtcgt tcatggtgac acttttagct ttgacatgat taagctcatc tcaattgatg
      481 ttatctaaag tcatttcaac tatctaagat gtggttgtga ttgggccatt ttgtgaaagc
      541 cagtacgcca gcgtcaatac actcccgtca attagttgca ccatgtccac aaaatcatat
      601 accagtagag ctgagactca tgcaagtccg gttgcatcga aacttttacg tttaatggat
      661 gaaaagaaga ccaatttgtg tgcttctctt gacgttcgtt cgactgatga gctattgaaa
      721 ctagttgaaa cgttgggtcc atacatttgc cttttgaaaa cacacgttga tatcttggat
      781 gatttcagtt atgagggtac tgtcgttcca ttgaaagcat tggcagagaa atacaagttc
      841 ttgatatttg aggacagaaa attcgccgat atcggtaaca cagtcaaatt acaatataca
      901 tcgggcgttt accgtatcgc agaatggtct gatatcacca acgcccacgg ggttactggt
      961 gctggtattg ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga accaagggga
     1021 ttattgatgc ttgctgaatt gtcttccaag ggttctctag cacacggtga atatactaag
     1081 ggtaccgttg atattgcaaa gagtgataaa gatttcgtta ttgggttcat tgctcagaac
     1141 gatatgggag gaagagaaga agggtttgat tggctaatca tgaccccagg tgtaggttta
     1201 gacgacaaag gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt tgtaagtggt
     1261 ggatcagata tcatcattgt tggcagagga cttttcgcca agggtagaga tcctaaggtt
     1321 gaaggtgaaa gatacagaaa tgctggatgg gaagcgtacc aaaagagaat cagcgctccc
     1381 cattaattat acaggaaact taatagaaca aatcacatat ttaatctaat agccacctgc
     1441 attggcacgg tgcaacacta cttcaacttc atcttacaaa aagatcacgt gatctgttgt
     1501 attgaactga aaattttttg tttgcttctc tctctctctt tcattatgtg agatttaaaa
     1561 accagaaact acatcatcga aaaagagttt taaaccatta caaccattgc gataagccct
     1621 ctcaaacttc ctccagactg tttgtcatcc aattggtaag attaattatc atataccccg
     1681 gataagaaaa gaagcaagca cacaatggat caacttaatg gtaaggaaca acaagcggcc
     1741 gccaccgcgg tggagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc
     1801 cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct
     1861 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1921 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1981 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2041 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2101 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2161 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2221 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2281 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2341 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2401 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2461 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2521 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2581 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2641 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2701 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2761 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2821 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2881 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2941 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3001 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3061 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3121 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3181 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3241 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3301 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3361 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3421 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3481 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3541 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3601 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3661 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3721 cctttttcaa taacctggct tatcgaaatt aatacgactc actataggga gaccggcaga
     3781 tccgcggccg cataggccac tagtggatct gatatcatcg atgaattcga gctcgttttc
     3841 gacactggat ggcggcgtta gtatcgaatc gacagcagta tagcgaccag cattcacata
     3901 cgattgacgc atgatattac tttctgcgca cttaacttcg catctgggca gatgatgtcg
     3961 aggcgaaaaa aaatataaat cacgctaaca tttgattaaa atagaacaac tacaatataa
     4021 aaaaactata caaatgacaa gttcttgaaa acaagaatct ttttattgtc agtactgatt
     4081 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     4141 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     4201 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     4261 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     4321 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     4381 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     4441 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     4501 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     4561 cttctaatac ctggaatgct gttttgccgg ggatcgcagt ggtgagtaac catgcatcat
     4621 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     4681 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     4741 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     4801 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     4861 tcgaaacgtg agtcttttcc ttacccatgg ttgtttatgt tcggatgtga tgtgagaact
     4921 gtatcctagc aagattttaa aaggaagtat atgaaagaag aacctcagtg gcaaatccta
     4981 accttttata tttctctaca ggggcgcggc gtggggacaa ttcaacgcgt ctgtgagggg
     5041 agcgtttccc tgctcgcagg tctgcagcga ggagccgtaa tttttgcttc gcgccgtgcg
     5101 gccatcaaaa tgtatggatg caaatgatta tacatgggga tgtatgggct aaatgtacgg
     5161 gcgacagtca catcatgccc ctgagctgcg cacgtcaaga ctgtcaagga gggtattctg
     5221 ggcctccatg tcgctggccg ggtgacccgg cggggacgag gcaagctaaa cagatctggc
     5281 gcgccttaat taacccggg
//
