LOCUS       sequence-11343-e        2868 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 2868)
  AUTHORS   Aoyama M, Agari K, Sun-Wada GH, Futai M, Wada Y
  TITLE     Simple and straightforward construction of a mouse gene targeting 
            vector using in vitro transposition reactions.
  JOURNAL   Nucleic Acids Res. 2005  . 33(5):e52.
  PUBMED    15784610
REFERENCE   2  (bases 1 to 2868)
  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: "Nucleic 
            Acids Res. 2005  . 33(5):e52."
COMMENT     Sequence Label: pMODloxZeo3DAmp
FEATURES             Location/Qualifiers
     source          1..2868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    309..327
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     protein_bind    complement(343..376)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    387..508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(398..417)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     452..471
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(572..946)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      943..952
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(956..974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(966..1013)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    complement(1079..1112)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1129..1147
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(1213..1230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1384..1972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1464..1483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct gtctcttata cacatctcaa ccatcatcga tcataacttc gtatagcata
      361 cattatacga agttatccag acatgataag atacattgat gagtttggac aaaccacaac
      421 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      481 aaccattata agctgcaata aacaagtttc gaggtcgacc cccctcggac ccgtcgggcc
      541 gcgtcggacc ggcggtgttg gtcggcgtcg gtcagtcctg ctcctcggcc acgaagtgca
      601 cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc tcgccgatct
      661 cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc gaccactcgg
      721 cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc ggcaccacct
      781 ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt
      841 cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc gctccggcga
      901 cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt ggccatggtg gccctcctat
      961 agtgagtcgt attatactat gccgatatac tatgccgatg attaattgtc aacacgtgcc
     1021 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     1081 aacttcgtat agcatacatt atacgaagtt atgatagctt cagggttgag atgtgtataa
     1141 gagacagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1201 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1261 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1321 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1381 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1441 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1501 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1561 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1621 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1681 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1741 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1801 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     1861 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1921 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1981 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2041 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2281 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtatta
     2701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2821 aaccattatt atcatgacat taacctataa aaataggcgt atcacgag
//
