LOCUS       sequence-323845-        2451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lactate-inducible expression of supP(opal), preceded by a hammerhead
            ribozyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2451)
  AUTHORS   Ho JML, Miller CA, Parks SE, Mattia JR, Bennett MR
  TITLE     A suppressor tRNA-mediated feedforward loop eliminates leaky gene 
            expression in bacteria.
  JOURNAL   Nucleic Acids Res. 2021 Mar 18;49(5):e25. doi: 10.1093/nar/gkaa1179.
  PUBMED    33290521
REFERENCE   2  (bases 1 to 2451)
  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: "Nucleic 
            Acids Res."; date: "2021-03-18"; volume: "49(5):e25. doi"; pages: " 
            10.1093/nar/gkaa1179"
COMMENT     Sequence Label: pJH773_pTech_PlldR-HHRz-supP(opal)
FEATURES             Location/Qualifiers
     source          1..2451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1127..1132,1133..1149,1150..1155))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      1267..1812
                     /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     1713..1732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1929..1946
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(2235..2451,1..443)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2301..2320)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
       61 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
      121 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggcgaa aacctggcct
      181 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
      241 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
      301 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
      361 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
      421 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
      481 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg aaagcaaatt
      541 cgacccgggc ggccgcaacc cagcgttcga tgcttctttg agcgaacgat caaaaataag
      601 tgccttccca tcaattggcc ctaccaattc ttcgcttatc tgacctctgg ttcacaattt
      661 cccaattaaa actcacatca atgttgccaa tacataacat ttagttaacc attcattgtc
      721 attatcccta cacaacacaa ttggcagtgc cacttgctat gagccgctga tgagtccgtg
      781 aggacgaaac ggtacccggt accgtcgccg aagtggcgaa atcggtagac gcagttgatt
      841 tcaaatcaac cgtagaaata cgtgccggtt cgagtccggc cttcggcacc aaattcgaaa
      901 agcctgctca acgagcaggc ttttttgcat ggcgttaccc aacttaatcg ccttgcagca
      961 catccccctt tcgccagatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     1021 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     1081 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     1141 atgataagct gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc
     1201 tacatttgaa gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg
     1261 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     1321 cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg
     1381 gcttggagga gcgcagccac caaaacttgt cctttcagtt tagccttaac cggcgcatga
     1441 cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat
     1501 gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa
     1561 cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca
     1621 ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca
     1681 ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct
     1741 gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg
     1801 agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg
     1861 catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga
     1921 ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct
     1981 gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc
     2041 agtgccaaca tagtaagcca gtatacactc cgctagcgct gatgtccggc ggtgcttttg
     2101 ccgttacgca ccaccccgtc agtagctgaa caggagggac agtaagaggt tccaactttc
     2161 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     2221 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     2281 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     2341 ccgttcagct ggacattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     2401 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa t
//
