LOCUS       sequence-12606-e        3246 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 3246)
  TITLE     latest plasmids
REFERENCE   2  (bases 1 to 3246)
  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: BirA in pENTR1A (w860-1)
FEATURES             Location/Qualifiers
     source          1..3246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /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             494..1456
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             494..1456
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     protein_bind    complement(1475..1574)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1686..1705)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1697..2506
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1764..1783)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2596..3184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3085..3104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt 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 gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccgaa ttcatgaagg ataacaccgt gccactgaaa ttgattgccc tgttagcgaa
      541 cggtgaattt cactctggcg agcagttggg tgaaacgctg ggaatgagcc gggcggctat
      601 taataaacac attcagacac tgcgtgactg gggcgttgat gtctttaccg ttccgggtaa
      661 aggatacagc ctgcctgagc ctatccagtt acttaatgct aaacagatat tgggtcagct
      721 ggatggcggt agtgtagccg tgctgccagt gattgactcc acgaatcagt accttcttga
      781 tcgtatcgga gagcttaaat cgggcgatgc ttgcattgca gaataccagc aggctggccg
      841 tggtcgccgg ggtcggaaat ggttttcgcc ttttggcgca aacttatatt tgtcgatgtt
      901 ctggcgtctg gaacaaggcc cggcggcggc gattggttta agtctggtta tcggtatcgt
      961 gatggcggaa gtattacgca agctgggtgc agataaagtt cgtgttaaat ggcctaatga
     1021 cctctatctg caggatcgca agctggcagg cattctggtg gagctgactg gcaaaactgg
     1081 cgatgcggcg caaatagtca ttggagccgg gatcaacatg gcaatgcgcc gtgttgaaga
     1141 gagtgtcgtt aatcaggggt ggatcacgct gcaggaagcg gggatcaatc tcgatcgtaa
     1201 tacgttggcg gccatgctaa tacgtgaatt acgtgctgcg ttggaactct tcgaacaaga
     1261 aggattggca ccttatctgt cgcgctggga aaagctggat aattttatta atcgcccagt
     1321 gaaacttatc attggtgata aagaaatatt tggcatttca cgcggaatag acaaacaggg
     1381 ggctttatta cttgagcagg atggaataat aaaaccctgg atgggcggtg aaatatccct
     1441 gcgtagtgca gaaaaaggta agcttgatat ctagacccag ctttcttgta caaagttggc
     1501 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1561 aaaatcatta tttgccatcc agctgcagct ctggcccgtg tctcaaaatc tctgatgtta
     1621 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     1681 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     1741 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     1801 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     1861 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     1921 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     1981 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2041 tgaaaatatt gttgatgcgc tggcagtgtc cctgcgccgg ttgcattcga ttcctgtttg
     2101 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2161 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2221 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2281 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2341 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2401 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2461 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2521 ttggttgtaa cattattcag attgggcccc gttccactga gcgtcagacc cggtagaaaa
     2581 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2641 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2701 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2761 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2821 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2881 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2941 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3001 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3061 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3121 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3181 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3241 catgtt
//
