LOCUS       sequence-366941-        3853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR with nt. 1-31 deleted, 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 3853)
  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 3853)
  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: pRAR039
FEATURES             Location/Qualifiers
     source          1..3853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(616..618,619..1332)
                     /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             616..1329
                     /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(646..674)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1269..1289
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1352..1381
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1397..1414
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1470..1487)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1640..1686
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1839..2498
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1905..1924)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2519..2613
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3020..3037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3154..3699)
                     /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(3234..3253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctttacact ttatgcttcc ggctgaattc taaagatctc ggcaaggcat gaaacagcac
       61 atcgccaatc ggatgatgat agtaatctgc cgcccatagc agcaatcgcc agacggagtg
      121 agtaaacacc ggctcactat ccagcacttc gactaccgct tttagctcat tgagcggcag
      181 ttcgctggca tcgctaactg ataccacaat cccgatgcgc tcctgctgtt tgccaaacgg
      241 cacgcgcacg cgacacccag ctttaaccgt catgccttct ggcagcagat agtcaaaggt
      301 acgaggaagc ggaacgggca aggcaacgtg ggcaacgggc atagcatcat cctgacttga
      361 aattcggtgg gttagtatac acattgccgt agaagagtgc ggatcagttt gcatacgctg
      421 gttaatttct gtatgatttc tatactttta acatcgcgtg gtgtctggcg ttagggctgg
      481 aagagcgacg cggccttaaa ccgaggtttt cccatgaaaa aagatattca cccgaaatac
      541 gaagaaatta ctgctagctg ctcttgcggt aacgtaatga aaatccgctc caccgttggt
      601 catggtggtg gtggtatgag caaaggagaa gaacttttca ctggagttgt cccaattctt
      661 gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt
      721 gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
      781 ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg
      841 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
      901 cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa
      961 ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac
     1021 attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac
     1081 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc
     1141 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     1201 ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt
     1261 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
     1321 ctctacaaat aaggatctga agcttgggcc cgaacaaaaa ctcatctcag aagaggatct
     1381 gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttggc
     1441 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     1501 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1561 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     1621 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1681 ttttatctgt tgtttgtcgg tgaactggat ccttactcga gtctagactg cagttgatcg
     1741 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     1801 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1861 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1921 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1981 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     2041 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     2101 ttaacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     2161 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     2221 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     2281 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     2341 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     2401 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     2461 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga
     2521 ctcctgttga tagatccagt aatgacctaa gaactccatc tggatttgtt cagaacgctc
     2581 ggttgccgcc gggcgttttt tattggtgag aatccaagcc tccgatcaac gtctcatttt
     2641 cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct
     2701 gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc
     2761 aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat
     2821 cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
     2881 acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     2941 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     3001 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     3061 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     3121 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     3181 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     3241 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     3301 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     3361 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     3421 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     3481 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     3541 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     3601 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     3661 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     3721 ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     3781 cccatacgat ataagttgta attctcatgt ttgacagctt atcatcgata agcttccgat
     3841 ggcgcgccga gag
//
