LOCUS       sequence-226864-        3100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal Tev-Avi tag Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3100)
  AUTHORS   Diebold-Durand ML, Burmann F, Gruber S
  TITLE     High-Throughput Allelic Replacement Screening in Bacillus subtilis.
  JOURNAL   Methods Mol Biol. 2019;2004:49-61. doi: 10.1007/978-1-4939-9520-2_5.
  PUBMED    31147909
REFERENCE   2  (bases 1 to 3100)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "2004:49-61. doi"; pages: " 
            10.1007/978-1-4939-9520-2_5"
COMMENT     Sequence Label: pJet1.2 Tev-Avi-tag
FEATURES             Location/Qualifiers
     source          1..3100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(395..439)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(452..472)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        complement(join(932..938,939..956,957..962))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    977..998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1118..1135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1289..1877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1369..1388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2048..2839,2840..2908))
                     /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     2671..2690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2909..3013)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3081..3099
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga ttcattgggt ctcacttatc tagtttcatg ccattcaatt ttttgtgctt
      421 caaaaatatc attcaggccg cctgcgctag caccctgaaa atacaaattc tctccgctgc
      481 cgcttgagac ctgtaacatc tttctagaag atctcctaca atattctcag ctgccatgga
      541 aaatcgatgt tcttctttta ttctctcaag attttcaggc tgtatattaa aacttatatt
      601 aagaactatg ctaaccacct catcaggaac cgttgtaggt ggcgtgggtt ttcttggcaa
      661 tcgactctca tgaaaactac gagctaaata ttcaatatgt tcctcttgac caactttatt
      721 ctgcattttt tttgaacgag gtttagagca agcttcagga aactgagaca ggaattttat
      781 taaaaattta aattttgaag aaagttcagg gttaatagca tccatttttt gctttgcaag
      841 ttcctcagca ttcttaacaa aagacgtctc ttttgacatg tttaaagttt aaacctcctg
      901 tgtgaaatta ttatccgctc ataattccac acattatacg agccggaagc ataaagtgta
      961 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgccaa
     1021 ttgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1081 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1141 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1981 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2041 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2101 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2161 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2221 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2281 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2341 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2401 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2461 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2521 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2581 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2641 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2701 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2761 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2821 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2881 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2941 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3001 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3061 tcatgacatt aacctataaa aataggcgta tcacgaggcc
//
