LOCUS       sequence-92127-e        3104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for amplification of a selection cassette that 
            includes two transcription terminator elements, an I-SceI site and a
            kanamycin resistance gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3104)
  AUTHORS   Kim J, Webb AM, Kershner JP, Blaskowski S, Copley SD
  TITLE     A versatile and highly efficient method for scarless genome editing 
            in Escherichia coli and Salmonella enterica.
  JOURNAL   BMC Biotechnol. 2014 Sep 25;14(1):84.
  PUBMED    25255806
REFERENCE   2  (bases 1 to 3104)
  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: "BMC 
            Biotechnol."; date: "2014-09-25"; volume: "14(1)"; pages: "84"
COMMENT     Sequence Label: pT2SK
FEATURES             Location/Qualifiers
     source          1..3104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(32..823,824..892))
                     /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     655..674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(893..997)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1065..1083
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1462..1481)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1473..2288
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1473..2288
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1546..1565)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2377..2965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2457..2476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
       61 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
      121 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      181 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      241 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      301 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      361 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      421 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      481 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      541 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      601 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      661 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      721 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      781 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      841 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      901 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      961 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1021 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1081 ggcccttatc tcaagagtgg cagcggttct gttaagtaac tgaacccaat gtcgttagtg
     1141 acgcttaccc gcaaaaaacc ccgcttcggc ggggtttttt cgctcttaag aggtcactga
     1201 cctaacaaaa aaaaaacccc gcccctgaca gggcggggtt ttttttggtc ttgagtggca
     1261 gagtcagtta tcgcgagcag tatgtaagta gatcctcagt gtcagctagg gataacaggg
     1321 taatctgatc cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc
     1381 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     1441 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     1501 gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1561 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     1621 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1681 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1741 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     1801 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1861 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     1921 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     1981 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     2041 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2101 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2161 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2221 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     2281 ttttctaagc agggcggggc gtaacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atct
//
