LOCUS       sequence-374745-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR G79U 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: pRAR106
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(636..638,639..1352)
                     /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             636..1349
                     /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(666..694)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1289..1309
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1372..1401
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1417..1434
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1490..1507)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1660..1706
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1859..2518
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1925..1944)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2539..2633
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3040..3057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3174..3719)
                     /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(3254..3273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatgcttccg gctgaattct aaagatctcg gcaaggcatg aaacagcaca tcgccaatcg
       61 gatgatgata gtaatctgcc gcccatagca gcaatcgcca gacggagtga gtaaacaccg
      121 gctcactatc cagcacttcg actaccgctt ttagctcatt gagcggcagt tcgctggcat
      181 cgctaactga taccacaatc ccgatgcgct cctgctgttt gccaaacggc acgcgcacgc
      241 gacacccagc tttaaccgtc atgccttctg gcagcagata gtcaaaggta cgaggaagcg
      301 gaacgggcaa ggcaacgtgg gcaacgggca tagcatcatc ctgacttgaa attcggtggg
      361 ttagtataca cattgccgta gaagagtgcg gatcagtttg catacgctgg ttaatttctg
      421 tatgatttcg cgccttcgta cgaaatgatc gtattgaagc tatactttta acatcgcgtg
      481 gtgtctggcg ttagggctgg aagagctacg cggccttaaa ccgaggtttt cccatgaaaa
      541 aagatattca cccgaaatac gaagaaatta ctgctagctg ctcttgcggt aacgtaatga
      601 aaatccgctc caccgttggt catggtggtg gtggtatgag caaaggagaa gaacttttca
      661 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      721 tccgtggaga gggtgaaggt gatgctacaa acggaaaact cacccttaaa tttatttgca
      781 ctactggaaa actacctgtt ccgtggccaa cacttgtcac tactctgacc tatggtgttc
      841 aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc
      901 ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggacc tacaagacgc
      961 gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg
     1021 attttaaaga agatggaaac attcttggac acaaactcga gtacaacttt aactcacaca
     1081 atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc
     1141 acaacgttga agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg
     1201 gcgatggccc tgtcctttta ccagacaacc attacctgtc gacacaatct gtcctttcga
     1261 aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga
     1321 ttacacatgg catggatgag ctctacaaat aaggatctga agcttgggcc cgaacaaaaa
     1381 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt
     1441 taaacggtct ccagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     1501 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     1561 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     1621 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     1681 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaactggat ccttactcga
     1741 gtctagactg cagttgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag
     1801 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
     1861 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     1921 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     1981 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     2041 cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga
     2101 gctggtgata tgggatagtg ttaacccttg ttacaccgtt ttccatgagc aaactgaaac
     2161 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     2221 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     2281 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     2341 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     2401 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     2461 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
     2521 tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctaa gaactccatc
     2581 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagcc
     2641 tccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
     2701 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa
     2761 gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct
     2821 ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt
     2881 aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg
     2941 gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt
     3001 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
     3061 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
     3121 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
     3181 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
     3241 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
     3301 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
     3361 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
     3421 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
     3481 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
     3541 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
     3601 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
     3661 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
     3721 aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa
     3781 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt ttgacagctt
     3841 atcatcgata agcttccgat ggcgcgccga gaggctttac actt
//
