LOCUS       sequence-64252-e        3177 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 3177)
  TITLE     Murray Lab pBEST plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3177)
  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            "Unpublished"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-pTar-UTR1-TetR-T500
FEATURES             Location/Qualifiers
     source          1..3177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             360..382
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             389..1012
                     /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(462..481)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1189..1206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1360..1948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1440..1459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2119..2910,2911..2979))
                     /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     2742..2761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2980..3084)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3152..3170
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcttcaagaa ttctggcgaa tcctctgacc agccagaaaa cgacctttct gtggtgaaac
       61 cggatgctgc aattcagagc ggcagcaagt gggggacagc agaagacctg accgccgcag
      121 agtggatgtt tgacatggtg aagactatcg caccatcagc cagaaaaccg aattttgctg
      181 ggtgggctaa cgatatccgc ctgatgcgtg aacgtgacgg acgtaaccac cgcgacatgt
      241 gtgtgctgtt ccgctgggca tgccaggaca acttctggtc cggtaacgtg ctgagcccgg
      301 ccaagcttca ataaagtttc ccccctcctt gccgataacg agatcaagct agcaataatt
      361 ttgtttaact ttaagaagga gatataccat gtcaagatta gataaaagta aagtgattaa
      421 cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc
      481 ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt
      541 gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt gccctttaga
      601 aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact
      661 aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga
      721 aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt
      781 atatgcactc agcgctgtgg ggcattttac tttaggttgc gtattggaag atcaagagca
      841 tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg
      901 acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga
      961 attgatcata tgcggattag aaaaacaact taaatgtgaa agtgggtctt aactcgagca
     1021 aagcccgccg aaaggcgggc ttttctgtgt cgaccgatgc ccttgagagc cttcaaccca
     1081 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     1141 tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga
     1201 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1261 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     1561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1801 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2101 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2161 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2221 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2281 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2341 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2401 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2461 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2521 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2581 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2641 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2701 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2761 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2821 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2881 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2941 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3001 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3061 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3121 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcg
//
