LOCUS       sequence-366943-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR with nt. 56-61 and 68-73 
            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: pRAR052
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(641..643,644..1357)
                     /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             641..1354
                     /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(671..699)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1294..1314
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1377..1406
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1422..1439
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1495..1512)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1665..1711
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1864..2523
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1930..1949)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2544..2638
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3045..3062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3179..3724)
                     /direction=LEFT
                     /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     complement(3259..3278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cactttatgc ttccggctga attctaaaga tctcggcaag gcatgaaaca gcacatcgcc
       61 aatcggatga tgatagtaat ctgccgccca tagcagcaat cgccagacgg agtgagtaaa
      121 caccggctca ctatccagca cttcgactac cgcttttagc tcattgagcg gcagttcgct
      181 ggcatcgcta actgatacca caatcccgat gcgctcctgc tgtttgccaa acggcacgcg
      241 cacgcgacac ccagctttaa ccgtcatgcc ttctggcagc agatagtcaa aggtacgagg
      301 aagcggaacg ggcaaggcaa cgtgggcaac gggcatagca tcatcctgac ttgaaattcg
      361 gtgggttagt atacacattg ccgtagaaga gtgcggatca gtttgcatac gctggttaat
      421 ttctgtatga tttcgcgcct tcgtacgaaa tgatcgtatt gaagctatac ttttaacatc
      481 gcgtggtgtg acgcgttagg gtgtcaagag cgacgcggcc ttaaaccgag gttttcccat
      541 gaaaaaagat attcacccga aatacgaaga aattactgct agctgctctt gcggtaacgt
      601 aatgaaaatc cgctccaccg ttggtcatgg tggtggtggt atgagcaaag gagaagaact
      661 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
      721 ttctgtccgt ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat
      781 ttgcactact ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg
      841 tgttcaatgc ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc
      901 catgcccgaa ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa
      961 gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg
     1021 tattgatttt aaagaagatg gaaacattct tggacacaaa ctcgagtaca actttaactc
     1081 acacaatgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat
     1141 tcgccacaac gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc
     1201 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct
     1261 ttcgaaagat cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc
     1321 tgggattaca catggcatgg atgagctcta caaataagga tctgaagctt gggcccgaac
     1381 aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt
     1441 gagtttaaac ggtctccagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     1501 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     1561 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     1621 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     1681 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac tggatcctta
     1741 ctcgagtcta gactgcagtt gatcgggcac gtaagaggtt ccaactttca ccataatgaa
     1801 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
     1861 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     1921 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     1981 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     2041 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
     2101 ggtgagctgg tgatatggga tagtgttaac ccttgttaca ccgttttcca tgagcaaact
     2161 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     2221 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     2281 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     2341 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     2401 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     2461 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     2521 taatttgata tcgagctcgc ttggactcct gttgatagat ccagtaatga cctaagaact
     2581 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     2641 aagcctccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac
     2701 agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc
     2761 gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag
     2821 gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg
     2881 tgcgtaacgg caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta
     2941 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     3001 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     3061 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     3121 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     3181 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     3241 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
     3301 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     3361 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     3421 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3481 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     3541 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     3601 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     3661 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     3721 tcaagaagat catcttatta atcagataaa atatttctag atttcagtgc aatttatctc
     3781 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgtttgac
     3841 agcttatcat cgataagctt ccgatggcgc gccgagaggc ttta
//
