LOCUS       sequence-92124-e        2946 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
            chloramphenicol resistance gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2946)
  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 2946)
  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: pT2SC
FEATURES             Location/Qualifiers
     source          1..2946
                     /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"
     promoter        1384..1486
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1487..2146
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1553..1572)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(2219..2807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2299..2318)
                     /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 taatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaga
     1381 cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     1441 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     1501 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1561 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     1621 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     1681 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     1741 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     1801 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     1861 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     1921 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     1981 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     2041 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     2101 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaacgca ggaaagaaca
     2161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2941 caatct
//
