LOCUS       sequence-496787-        2951 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  10xTetO Sponge Plasmid with BsaI sites flanking sponge casette for 
            Golden Gate Assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2951)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 2951)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEM413
FEATURES             Location/Qualifiers
     source          1..2951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    39..57
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    86..104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    135..153
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    167..185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    209..227
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    255..273
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    297..315
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    345..363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    394..412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    436..454
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      489..560
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      576..603
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(691..1506)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(691..1506)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1414..1433
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1498..1517
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1967..1984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2101..2645)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2180..2199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgagcggcc gcttctagag atcgatttta gagctgcctc cctatcagtg atagagattg
       61 ctacaactgc aaggtagtcc cggtatccct atcagtgata gagactgata gctacgagtc
      121 tgagatccta cttgtcccta tcagtgatag agacagtacc gagttttccc tatcagtgat
      181 agagagctca tactgcgaca gagtcttttc cctatcagtg atagagatag gctcctttgt
      241 caaaagtgat cgcttcccta tcagtgatag agacttcacg ggataatgtg ttctactccc
      301 tatcagtgat agagagctta agtcaatcca atgcgttcag gtgctcccta tcagtgatag
      361 agaaactccg tcatcgtgga tgtcaatact gactccctat cagtgataga gattgatttc
      421 accgctggcg tcaattccct atcagtgata gagacttagg tgtccctaga attactagag
      481 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
      541 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
      601 gcgtttatat atgatcgaga ccggtaatct cgagtcccgt caagtcagcg taatgctctg
      661 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
      721 ctgcaattta ttcatatcag gattatcgat accatatttt tgaaaaagcc gtttctgtaa
      781 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
      841 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
      901 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
      961 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1021 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1081 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1141 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     1201 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1261 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1321 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1381 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     1441 atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg
     1501 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     1561 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     1621 aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga
     1681 ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct
     1741 catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga
     1801 gtaagcaaca ccttcttcac gaggcagacc tcagcgctag cggagtgtat actggcttac
     1861 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     1921 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     1981 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     2041 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     2101 tttccatagg ctccgccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     2161 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     2221 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     2281 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     2341 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2401 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     2461 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     2521 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcaaaaaac
     2581 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     2641 ctcaagaaga tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2941 tgatggtctc g
//
