LOCUS       sequence-220459-        3221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3221)
  TITLE     latest plasmids
REFERENCE   2  (bases 1 to 3221)
  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"
COMMENT     Sequence Label: pENTR4-HaloTag (w876-1)
FEATURES             Location/Qualifiers
     source          1..3221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      106..149
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             463..1353
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1366..1386
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     protein_bind    complement(1450..1549)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1661..1680)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1672..2481
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1739..1758)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2571..3159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3060..3079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggcc
      121 cagtcttccg actgagcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatggcaga aatcggtact
      481 ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat
      541 gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac
      601 gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg
      661 atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc
      721 ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac
      781 tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt
      841 gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc
      901 gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac
      961 gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg
     1021 gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca
     1081 aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg
     1141 gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg
     1201 atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac
     1261 atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc
     1321 gcgcgctggc tgtcgacgct cgagatttcc ggcgagccaa ccactgagga tctgtacttt
     1381 cagagcgata acgcgaattc agtcgactgg atccggtacc gaattcgcgg ccgcactcga
     1441 gatatctaga cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     1501 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1561 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1621 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1681 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1741 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1801 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1861 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1921 cattttatcc gtactcctgg tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1981 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2041 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     2101 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2161 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     2221 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2281 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2341 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2401 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2461 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg
     2521 gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2701 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3181 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
