LOCUS       sequence-366954-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR U54C mutation, 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: pRAR102
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(643..645,646..1359)
                     /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             643..1356
                     /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(673..701)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1296..1316
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1379..1408
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1424..1441
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1497..1514)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1667..1713
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1866..2525
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFASFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1932..1951)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2546..2640
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3047..3064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3181..3726)
                     /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(3261..3280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacactttat gcttccggct gaattctaaa gatctcggca aggcatgaaa cagcacatcg
       61 ccaatcggat gatgatagta atctgccgcc catagcagca atcgccagac ggagtgagta
      121 aacaccggct cactatccag cacttcgact accgctttta gctcattgag cggcagttcg
      181 ctggcatcgc taactgatac cacaatcccg atgcgctcct gctgtttgcc aaacggcacg
      241 cgcacgcgac acccagcttt aaccgtcatg ccttctggca gcagatagtc aaaggtacga
      301 ggaagcggaa cgggcaaggc aacgtgggca acgggcatag catcatcctg acttgaaatt
      361 cggtgggtta gtatacacat tgccgtagaa gagtgcggat cagtttgcat acgctggtta
      421 atttctgtat gatttcgcgc cttcgtacga aatgatcgta ttgaagctat acttttaaca
      481 tcgcgtggcg tctggcgtta gggctggaag agcgacgcgg ccttaaaccg aggttttccc
      541 atgaaaaaag atattcaccc gaaatacgaa gaaattactg ctagctgctc ttgcggtaac
      601 gtaatgaaaa tccgctccac cgttggtcat ggtggtggtg gtatgagcaa aggagaagaa
      661 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      721 ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt
      781 atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat
      841 ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt
      901 gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac
      961 aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag
     1021 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac
     1081 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa
     1141 attcgccaca acgttgaaga tggttccgtt caactagcag accattatca acaaaatact
     1201 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc
     1261 ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct
     1321 gctgggatta cacatggcat ggatgagctc tacaaataag gatctgaagc ttgggcccga
     1381 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca
     1441 ttgagtttaa acggtctcca gcttggctgt tttggcggat gagagaagat tttcagcctg
     1501 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     1561 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     1621 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     1681 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga actggatcct
     1741 tactcgagtc tagactgcag ttgatcgggc acgtaagagg ttccaacttt caccataatg
     1801 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1861 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1921 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1981 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2041 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     2101 acggtgagct ggtgatatgg gatagtgtta acccttgtta caccgttttc catgagcaaa
     2161 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2221 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2281 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcgccagt tttgatttaa
     2341 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     2401 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     2461 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2521 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctaagaa
     2581 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     2641 ccaagcctcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     2701 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     2761 gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
     2821 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
     2881 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     2941 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     3001 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     3061 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     3121 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     3181 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     3241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     3301 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     3361 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     3421 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3481 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     3541 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     3601 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     3661 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     3721 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     3781 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     3841 acagcttatc atcgataagc ttccgatggc gcgccgagag gctt
//
