LOCUS       sequence-495595-        3411 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  20xTetO 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 3411)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 3411)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEM414
FEATURES             Location/Qualifiers
     source          1..3411
                     /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"
     protein_bind    499..517
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    546..564
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    595..613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    627..645
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    669..687
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    715..733
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    757..775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    805..823
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    854..872
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    896..914
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      949..1020
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1036..1063
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1151..1966)
                     /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(1151..1966)
                     /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     1874..1893
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1958..1977
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2427..2444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2561..3105)
                     /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(2640..2659)
                     /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 atcgatttta gagctgcctc cctatcagtg atagagattg ctacaactgc aaggtagtcc
      541 cggtatccct atcagtgata gagactgata gctacgagtc tgagatccta cttgtcccta
      601 tcagtgatag agacagtacc gagttttccc tatcagtgat agagagctca tactgcgaca
      661 gagtcttttc cctatcagtg atagagatag gctcctttgt caaaagtgat cgcttcccta
      721 tcagtgatag agacttcacg ggataatgtg ttctactccc tatcagtgat agagagctta
      781 agtcaatcca atgcgttcag gtgctcccta tcagtgatag agaaactccg tcatcgtgga
      841 tgtcaatact gactccctat cagtgataga gattgatttc accgctggcg tcaattccct
      901 atcagtgata gagacttagg tgtccctaga attactagag ccaggcatca aataaaacga
      961 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1021 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat atgatcgaga
     1081 ccggtaatct cgagtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa
     1141 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     1201 gattatcgat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1261 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1321 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1381 gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt
     1441 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1501 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     1561 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1621 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     1681 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1741 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1801 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1861 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1921 ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     1981 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     2041 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg
     2101 agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa
     2161 agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca
     2221 ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac
     2281 gaggcagacc tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg
     2341 tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat
     2401 atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     2461 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     2521 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     2581 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     2641 aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg
     2701 gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca
     2761 gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg
     2821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag
     2881 caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc
     2941 ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct
     3001 cggttcaaag agttggtagc tcagagaacc ttcaaaaaac cgccctgcaa ggcggttttt
     3061 tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt
     3121 aaggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3181 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3241 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3301 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3361 acgatacggg agggcttacc atctggcccc agtgctgcaa tgatggtctc g
//
