LOCUS       sequence-363327-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR with nt. 47-51 and 80-84 
            mutated, 90 nt of l31 coding region, glycine linker, sfGFP, trrnB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3884)
  AUTHORS   Rasmussen RA, Wang S, Camarillo JM, Sosnowski V, Cho BK, Goo YA, 
            Lucks JB, O'Halloran TV
  TITLE     Zur and zinc increase expression of E. coli ribosomal protein L31 
            through RNA-mediated repression of the repressor L31p.
  JOURNAL   Nucleic Acids Res. 2022 Dec 9;50(22):12739-12753. doi: 
            10.1093/nar/gkac1086.
  PUBMED    36533433
REFERENCE   2  (bases 1 to 3884)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2022-12-9- 9"; volume: "50"; issue: 
            "22"; pages: "12739-12753"
COMMENT     Sequence Label: pRAR051
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      157..702
                     /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     603..622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     819..836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1243..1337)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1358..2017)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1932..1951
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(2170..2216)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2369..2386
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2442..2459)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2475..2504)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(2524..3237,3238..3240))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(2527..3240)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2567..2587)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3182..3210
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gcctctcggc gcgccatcgg aagcttatcg atgataagct gtcaaacatg agaattacaa
       61 cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat
      121 ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt
      181 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
      241 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt
      301 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
      361 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
      421 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
      481 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
      541 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
      601 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
      661 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
      721 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
      781 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
      841 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
      901 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc
      961 cgctagcgct gatgtccggc ggtgcttttg ccgttacgca ccaccccgtc agtagctgaa
     1021 caggagggac agctgataga aacagaagcc actggagcac ctcaaaaaca ccatcataca
     1081 ctaaatcagt aagttggcag catcacccga cgcactttgc gccgaataaa tacctgtgac
     1141 ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg
     1201 ggccaacttt tggcgaaaat gagacgttga tcggaggctt ggattctcac caataaaaaa
     1261 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttcttagg tcattactgg
     1321 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     1381 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     1441 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     1501 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     1561 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     1621 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     1681 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1741 tgtaacaagg gttaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     1801 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     1861 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     1921 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     1981 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     2041 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     2101 aacctcttac gtgcccgatc aactgcagtc tagactcgag taaggatcca gttcaccgac
     2161 aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     2221 ctggcagttc cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt
     2281 cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt
     2341 ctgttttatc agaccgcttc tgcgttctga tttaatctgt atcaggctga aaatcttctc
     2401 tcatccgcca aaacagccaa gctggagacc gtttaaactc aatgatgatg atgatgatgg
     2461 tcgacggcgc tattcagatc ctcttctgag atgagttttt gttcgggccc aagcttcaga
     2521 tccttatttg tagagctcat ccatgccatg tgtaatccca gcagcagtta caaactcaag
     2581 aaggaccatg tggtcacgct tttcgttggg atctttcgaa aggacagatt gtgtcgacag
     2641 gtaatggttg tctggtaaaa ggacagggcc atcgccaatt ggagtatttt gttgataatg
     2701 gtctgctagt tgaacggaac catcttcaac gttgtggcga attttgaagt tagctttgat
     2761 tccattcttt tgtttgtctg ccgtgatgta tacattgtgt gagttaaagt tgtactcgag
     2821 tttgtgtcca agaatgtttc catcttcttt aaaatcaata ccctttaact cgatacgatt
     2881 aacaagggta tcaccttcaa acttgacttc agcacgcgtc ttgtaggtcc cgtcatcttt
     2941 gaaagatata gtgcgttcct gtacataacc ttcgggcatg gcactcttga aaaagtcatg
     3001 ccgtttcatg tgatccggat aacgggaaaa gcattgaaca ccataggtca gagtagtgac
     3061 aagtgttggc cacggaacag gtagttttcc agtagtgcaa ataaatttaa gggtgagttt
     3121 tccgtttgta gcatcacctt caccctctcc acggacagaa aatttgtgcc cattaacatc
     3181 accatctaat tcaacaagaa ttgggacaac tccagtgaaa agttcttctc ctttgctcat
     3241 accaccacca ccatgaccaa cggtggagcg gattttcatt acgttaccgc aagagcagct
     3301 agcagtaatt tcttcgtatt tcgggtgaat atcttttttc atgggaaaac ctcggtttaa
     3361 ggcgcgcacg ctcttccagc cctaacgcca gacacctgcg catgttaaaa gtatagcttc
     3421 aatacgatca tttcgtacga aggcgcgaaa tcatacagaa attaaccagc gtatgcaaac
     3481 tgatccgcac tcttctacgg caatgtgtat actaacccac cgaatttcaa gtcaggatga
     3541 tgctatgccc gttgcccacg ttgccttgcc cgttccgctt cctcgtacct ttgactatct
     3601 gctgccagaa ggcatgacgg ttaaagctgg gtgtcgcgtg cgcgtgccgt ttggcaaaca
     3661 gcaggagcgc atcgggattg tggtatcagt tagcgatgcc agcgaactgc cgctcaatga
     3721 gctaaaagcg gtagtcgaag tgctggatag tgagccggtg tttactcact ccgtctggcg
     3781 attgctgcta tgggcggcag attactatca tcatccgatt ggcgatgtgc tgtttcatgc
     3841 cttgccgaga tctttagaat tcagccggaa gcataaagtg taaa
//
