LOCUS       sequence-496745-        4331 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  40xTetO 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 4331)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 4331)
  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: pEM415
FEATURES             Location/Qualifiers
     source          1..4331
                     /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"
     protein_bind    959..977
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1006..1024
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1055..1073
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1087..1105
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1129..1147
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1175..1193
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1217..1235
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1265..1283
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1314..1332
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1356..1374
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1419..1437
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1466..1484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1515..1533
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1547..1565
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1589..1607
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1635..1653
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1677..1695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1725..1743
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1774..1792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1816..1834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1869..1940
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1956..1983
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2071..2886)
                     /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(2071..2886)
                     /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     2794..2813
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2878..2897
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3347..3364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3481..4025)
                     /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(3560..3579)
                     /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 atcgatttta gagctgcctc
      961 cctatcagtg atagagattg ctacaactgc aaggtagtcc cggtatccct atcagtgata
     1021 gagactgata gctacgagtc tgagatccta cttgtcccta tcagtgatag agacagtacc
     1081 gagttttccc tatcagtgat agagagctca tactgcgaca gagtcttttc cctatcagtg
     1141 atagagatag gctcctttgt caaaagtgat cgcttcccta tcagtgatag agacttcacg
     1201 ggataatgtg ttctactccc tatcagtgat agagagctta agtcaatcca atgcgttcag
     1261 gtgctcccta tcagtgatag agaaactccg tcatcgtgga tgtcaatact gactccctat
     1321 cagtgataga gattgatttc accgctggcg tcaattccct atcagtgata gagacttagg
     1381 tgtccctaga attactagag atcgatttta gagctgcctc cctatcagtg atagagattg
     1441 ctacaactgc aaggtagtcc cggtatccct atcagtgata gagactgata gctacgagtc
     1501 tgagatccta cttgtcccta tcagtgatag agacagtacc gagttttccc tatcagtgat
     1561 agagagctca tactgcgaca gagtcttttc cctatcagtg atagagatag gctcctttgt
     1621 caaaagtgat cgcttcccta tcagtgatag agacttcacg ggataatgtg ttctactccc
     1681 tatcagtgat agagagctta agtcaatcca atgcgttcag gtgctcccta tcagtgatag
     1741 agaaactccg tcatcgtgga tgtcaatact gactccctat cagtgataga gattgatttc
     1801 accgctggcg tcaattccct atcagtgata gagacttagg tgtccctaga attactagag
     1861 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1921 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     1981 gcgtttatat atgatcgaga ccggtaatct cgagtcccgt caagtcagcg taatgctctg
     2041 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     2101 ctgcaattta ttcatatcag gattatcgat accatatttt tgaaaaagcc gtttctgtaa
     2161 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     2221 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     2281 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
     2341 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     2401 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     2461 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     2521 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     2581 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     2641 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     2701 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     2761 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     2821 atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg
     2881 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     2941 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     3001 aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga
     3061 ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct
     3121 catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga
     3181 gtcagcaaca ccttcttcac gaggcagacc tcagcgctag cggagtgtat actggcttac
     3241 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     3301 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     3361 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     3421 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     3481 tttccatagg ctccgccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     3601 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     3661 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     3721 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3781 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     3841 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     3901 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcaaaaaac
     3961 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     4021 ctcaagaaga tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4081 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4141 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4201 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4261 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4321 tgatggtctc g
//
