LOCUS       sequence-226886-        3922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal Tev Halo-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3922)
  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 3922)
  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-Halo tag
FEATURES             Location/Qualifiers
     source          1..3922
                     /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(383..1267)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             complement(1280..1300)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        complement(join(1754..1760,1761..1778,1779..1784))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1799..1820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1940..1957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2111..2699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2191..2210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2870..3661,3662..3730))
                     /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     3493..3512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3731..3835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3903..3921
                     /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 tggtctcgct taaccggaaa tctccagagt agacagccag cgcgcgatct
      421 cgctgccgat caggtccggg ttgtcttctt gcagcagatt cagacccggg ccgatgtcca
      481 cagccttgca gttaggcagg cttttggcca ggcgagcggc ttcggccggt gggatcagaa
      541 cgcctggggt gccccagaac agcagcttcg ggacagggga ctggtgcagc cagtccatgt
      601 attcttcgac cagcgcgacg atgttcgctg gctcaccggc gattggcagc tcgtttggga
      661 agcgccacag tggctcgcgg tcaacaggat tcaggaacgg ctcgcggtaa tggtccatct
      721 cgacttcagt cagcgggcgg acgacaccca tcggcagcgt accctcgata aaaacgttct
      781 gatcgatgat cagcttgcgg ccgacgtcgg tggtgcggaa ggcctggaag gtttcgcggg
      841 caaattctgg ccattcgtcc caggtcggga tagggcggat gaactccata aatgcaatac
      901 ctttgacgcg ctctggattg cgcttggccc agtggaaacc cagagcggag ccccagtcgt
      961 gaatgaccag gacgacctct tccagaccca gggcttcgat gaaggcatcc atgaagcgga
     1021 cgtggtcgtc gaagaaataa cccaggtctg gtttgtcgga tttgcccata ccgatcaggt
     1081 ctggagcaat gcagcgatgg gtcggtgcaa catgcgggat gatgttgcgc cacacgtagg
     1141 aggaggtcgg gttaccgtgc aggaacagca caggggtgcc atcgcgcgga ccaacatcga
     1201 cgtagtgcat gcgctcgccc aggacttcca cataatgggg gtcgaatgga aagccagtac
     1261 cgatttcgct tccgctagca ccctgaaaat acaaattctc tccgcttccg ctcgagacca
     1321 tctttctaga agatctccta caatattctc agctgccatg gaaaatcgat gttcttcttt
     1381 tattctctca agattttcag gctgtatatt aaaacttata ttaagaacta tgctaaccac
     1441 ctcatcagga accgttgtag gtggcgtggg ttttcttggc aatcgactct catgaaaact
     1501 acgagctaaa tattcaatat gttcctcttg accaacttta ttctgcattt tttttgaacg
     1561 aggtttagag caagcttcag gaaactgaga caggaatttt attaaaaatt taaattttga
     1621 agaaagttca gggttaatag catccatttt ttgctttgca agttcctcag cattcttaac
     1681 aaaagacgtc tcttttgaca tgtttaaagt ttaaacctcc tgtgtgaaat tattatccgc
     1741 tcataattcc acacattata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1801 gagtgagcta actcacatta attgcgttgc gctcactgcc aattgctttc cagtcgggaa
     1861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3841 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3901 aaaataggcg tatcacgagg cc
//
