LOCUS       sequence-274733-        3206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TetR ORF with stop codon between attL1-attL2 on Gateway entry 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3206)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3206)
  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: pENTR-TetR
FEATURES             Location/Qualifiers
     source          1..3206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             674..1294
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(747..766)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    complement(1331..1430)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1447..1466)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1448..1466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1471..1487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1471..1487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1589..1608)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1600..2409
                     /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
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1667..1686)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2556..3144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3045..3064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatgtcta gattagataa aagtaaagtg attaacagcg cattagagct
      721 gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt
      781 agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc
      841 cattgagatg ttagataggc accatactca cttttgccct ttagaagggg aaagctggca
      901 agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg
      961 agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca
     1021 attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc
     1081 tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga
     1141 agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt
     1201 atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg
     1261 attagaaaaa caacttaaat gtgaaagtgg gtccgcgtac agcggatccc gggaattcag
     1321 atcttattaa acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1381 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1441 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     1501 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1561 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1621 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1681 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     1741 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1801 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     1861 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     1921 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     1981 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2041 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2101 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2161 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2221 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2281 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2341 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2401 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     2461 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     2521 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2581 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2641 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2701 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2761 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2821 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2881 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2941 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3001 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3061 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3121 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3181 ttttgctggc cttttgctca catgtt
//
