LOCUS       sequence-64258-e        3433 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 3433)
  TITLE     Murray Lab pBEST plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3433)
  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            "Unpublished"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-p15a-pBAD-UTR1-TetR-T500
FEATURES             Location/Qualifiers
     source          1..3433
                     /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..659
                     /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"
     rep_origin      946..1490
                     /direction=RIGHT
                     /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     1391..1410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1607..1624
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(1857..2648,2649..2717))
                     /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     2480..2499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2718..2822)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2890..2908
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     3385..3404
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
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 gggtcttaac
      661 tcgagcaaag cccgccgaaa ggcgggcttt tctgtgtcga ccgatgccct tgagagcctt
      721 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
      781 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca tgataagctg tcaaacatga
      841 gaattacaac ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg
      901 cactgaaatc tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg
      961 tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa
     1021 ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc
     1081 aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa
     1141 atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa
     1201 gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt
     1261 ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc
     1321 ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg
     1381 gagccgccag gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat
     1441 ttgagcgtca gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg
     1501 ccgcggccct ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc
     1561 ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag
     1621 cgaggaagcg gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt
     1681 tctcctgcca catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt
     1741 atacactccg ctagggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2821 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2881 acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattctg gcgaatcctc
     2941 tgaccagcca gaaaacgacc tttctgtggt gaaaccggat gctgcaattc agagcggcag
     3001 caagtggggg acagcagaag acctgaccgc cgcagagtgg atgtttgaca tggtgaagac
     3061 tatcgcacca tcagccagaa aaccgaattt tgctgggtgg gctaacgata tccgcctgat
     3121 gcgtgaacgt gacggacgta accaccgcga catgtgtgtg ctgttccgct gggcatgcag
     3181 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
     3241 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     3301 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     3361 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc
     3421 ctacctggct agc
//
