LOCUS       sequence-92128-e        2770 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
            trimethoprim resistance gene. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2770)
  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 2770)
  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: pT2ST
FEATURES             Location/Qualifiers
     source          1..2770
                     /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"
     rep_origin      complement(2043..2631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2123..2142)
                     /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 taatcggata gacggcatgc acgatttgta ataacagagt gtcttgtatt tttaaagaaa
     1381 gtctatttaa tacaagtgat tatattaatt aacggtaagc atcagcgggt gacaaaacga
     1441 gcatgcttac taataaaatg ttaacctctg aggaagaatt gtgaaactat cactaatggt
     1501 agctatatcg aagaatggag ttatcgggaa tggccctgat attccatgga gtgccaaagg
     1561 tgaacagctc ctgtttaaag ctattaccta taaccaatgg ctgttggttg gacgcaagac
     1621 ttttgaatca atgggagcat tacccaaccg aaagtatgcg gtcgtaacac gttcaagttt
     1681 tacatctgac aatgaggacg tattgatctt tccatcaatt aaagatgctt taaccaacct
     1741 aaagaaaata acggatcatg tcattgtttc aggtggtggg gagatataca aaagcctgat
     1801 cgatcaagta gatacactac atatatctac aatagacatc gagccggaag gtgatgttta
     1861 ctttcctgaa atccccagca attttaggcc agtttttacc caagacttcg cctctaacat
     1921 aaattatagt taccaaatct ggcaaaaggg ttaagcaggg cggggcgtaa cgcaggaaag
     1981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2221 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2461 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2761 aaatcaatct
//
