LOCUS       sequence-64250-e        3166 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 3166)
  TITLE     Murray Lab pBEST plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3166)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBEST-PLlacO-1-UTR1-TetR-YbaQ-T500
FEATURES             Location/Qualifiers
     source          1..3166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             36..656
                     /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(109..128)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(859..876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1030..1618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1110..1129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1789..2580,2581..2649))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2412..2431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2650..2754)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2822..2840
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    3131..3147
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgtc tagattagat aaaagtaaag
       61 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
      121 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
      181 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
      241 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
      301 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
      361 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
      421 atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc
      481 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
      541 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
      601 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtctagaa
      661 gggaagaaag agcaaagaag gtagcataac tcgagcaaag cccgccgaaa ggcgggcttt
      721 tctgtgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
      781 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
      841 gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      901 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      961 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1021 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1081 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1141 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1201 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     1261 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1321 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1381 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1441 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     1501 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1561 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1621 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1681 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1741 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1801 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1861 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1921 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1981 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2041 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2101 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2161 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2221 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2281 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2341 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2401 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2461 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2521 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2581 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2641 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2701 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2761 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     2821 aaataggcgt atcacgaggc cctttcgtct tcaagaattc tggcgaatcc tctgaccagc
     2881 cagaaaacga cctttctgtg gtgaaaccgg atgctgcaat tcagagcggc agcaagtggg
     2941 ggacagcaga agacctgacc gccgcagagt ggatgtttga catggtgaag actatcgcac
     3001 catcagccag aaaaccgaat tttgctgggt gggctaacga tatccgcctg atgcgtgaac
     3061 gtgacggacg taaccaccgc gacatgtgtg tgctgttccg ctgcataaat gtgagcggat
     3121 aacattgaca ttgtgagcgg ataacaagat actgagcaca gctagc
//
