LOCUS       sequence-226900-        3991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal HA-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3991)
  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 3991)
  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 HA6-Tag
FEATURES             Location/Qualifiers
     source          1..3991
                     /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..484)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(491..580)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     promoter        complement(join(1046..1052,1053..1070,1071..1076))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     mobile_element  1089..1856
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     protein_bind    1868..1889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2009..2026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2180..2768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2260..2279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2939..3730,3731..3799))
                     /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     3562..3581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3800..3904)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3972..3990
                     /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 ctcgcttagc tagaagcgta atctggaacg tcatatggat
      421 aggatcctgc atagtccggg acgtcatagg gatagcccgc atagtcagga acatcgtatg
      481 ggtatctaga agcgtaatct ggaacgtcat atggatagga tcctgcatag tccgggacgt
      541 catagggata gcccgcatag tcaggaacat cgtatgggta tctagcagca gaaccgcttg
      601 agacctgtaa catctttcta gaagatctcc tacaatattc tcagctgcca tggaaaatcg
      661 atgttcttct tttattctct caagattttc aggctgtata ttaaaactta tattaagaac
      721 tatgctaacc acctcatcag gaaccgttgt aggtggcgtg ggttttcttg gcaatcgact
      781 ctcatgaaaa ctacgagcta aatattcaat atgttcctct tgaccaactt tattctgcat
      841 tttttttgaa cgaggtttag agcaagcttc aggaaactga gacaggaatt ttattaaaaa
      901 tttaaatttt gaagaaagtt cagggttaat agcatccatt ttttgctttg caagttcctc
      961 agcattctta acaaaagacg tctcttttga catgtttaaa gtttaaacct cctgtgtgaa
     1021 attattatcc gctcataatt ccacacatta tacgagccgg aagcataaag tgtaaagcct
     1081 ggggtgccgg taatgactcc aacttattga tagtgtttta tgttcagata atgcccgatg
     1141 actttgtcat gcagctccac cgattttgag aacgacagcg acttccgtcc cagccgtgcc
     1201 aggtgctgcc tcagattcag gttatgccgc tcaattcgct gcgtatatcg cttgctgatt
     1261 acgtgcagct ttcccttcag gcgggattca tacagcggcc agccatccgt catccatatc
     1321 accacgtcaa agggtgacag caggctcata agacgcccca gcgtcgccat agtgcgttca
     1381 ccgaatacgt gcgcaacaac cgtcttccgg agactgtcat acgcgtaaaa cagccagcgc
     1441 tggcgcgatt tagccccgac atagccccac tgttcgtcca tttccgcgca gacgatgacg
     1501 tcactgcccg gctgtatgcg cgaggttacc gactgcggcc tgagtttttt aagtgacgta
     1561 aaatcgtgtt gaggccaacg cccataatgc gggctgttgc ccggcatcca acgccattca
     1621 tggccatatc aatgattttc tggtgcgtac cgggttgaga agcggtgtaa gtgaactgca
     1681 gttgccatgt tttacggcag tgagagcaga gatagcgctg atgtccggcg gtgcttttgc
     1741 cgttacgcac caccccgtca gtagctgaac aggagggaca gctgatagaa acagaagcca
     1801 ctggagcacc tcaaaaacac catcatacac taaatcagta agttggcagc atcacctggg
     1861 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgcca attgctttcc
     1921 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1981 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3901 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     3961 taacctataa aaataggcgt atcacgaggc c
//
