LOCUS       sequence-363329-        3882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR G52 and G79 deletion, 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 3882)
  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 3882)
  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: pRAR105
FEATURES             Location/Qualifiers
     source          1..3882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(654..656,657..1370)
                     /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             654..1367
                     /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(684..712)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1307..1327
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1390..1419
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1435..1452
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1508..1525)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1678..1724
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1877..2536
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1943..1962)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2557..2651
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3058..3075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3192..3737)
                     /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(3272..3291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccgagagg ctttacactt tatgcttccg gctgaattct aaagatctcg gcaaggcatg
       61 aaacagcaca tcgccaatcg gatgatgata gtaatctgcc gcccatagca gcaatcgcca
      121 gacggagtga gtaaacaccg gctcactatc cagcacttcg actaccgctt ttagctcatt
      181 gagcggcagt tcgctggcat cgctaactga taccacaatc ccgatgcgct cctgctgttt
      241 gccaaacggc acgcgcacgc gacacccagc tttaaccgtc atgccttctg gcagcagata
      301 gtcaaaggta cgaggaagcg gaacgggcaa ggcaacgtgg gcaacgggca tagcatcatc
      361 ctgacttgaa attcggtggg ttagtataca cattgccgta gaagagtgcg gatcagtttg
      421 catacgctgg ttaatttctg tatgatttcg cgccttcgta cgaaatgatc gtattgaagc
      481 tatactttta acatcgcgtg tgtctggcgt tagggctgga agagcacgcg gccttaaacc
      541 gaggttttcc catgaaaaaa gatattcacc cgaaatacga agaaattact gctagctgct
      601 cttgcggtaa cgtaatgaaa atccgctcca ccgttggtca tggtggtggt ggtatgagca
      661 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
      721 atgggcacaa attttctgtc cgtggagagg gtgaaggtga tgctacaaac ggaaaactca
      781 cccttaaatt tatttgcact actggaaaac tacctgttcc gtggccaaca cttgtcacta
      841 ctctgaccta tggtgttcaa tgcttttccc gttatccgga tcacatgaaa cggcatgact
      901 ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct ttcaaagatg
      961 acgggaccta caagacgcgt gctgaagtca agtttgaagg tgataccctt gttaatcgta
     1021 tcgagttaaa gggtattgat tttaaagaag atggaaacat tcttggacac aaactcgagt
     1081 acaactttaa ctcacacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag
     1141 ctaacttcaa aattcgccac aacgttgaag atggttccgt tcaactagca gaccattatc
     1201 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcga
     1261 cacaatctgt cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc cttcttgagt
     1321 ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaataa ggatctgaag
     1381 cttgggcccg aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat
     1441 catcatcatc attgagttta aacggtctcc agcttggctg ttttggcgga tgagagaaga
     1501 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     1561 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     1621 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     1681 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1741 aactggatcc ttactcgagt ctagactgca gttgatcggg cacgtaagag gttccaactt
     1801 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     1861 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     1921 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     1981 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     2041 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat
     2101 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt aacccttgtt acaccgtttt
     2161 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     2221 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     2281 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     2341 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     2401 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     2461 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     2521 gcagggcggg gcgtaatttg atatcgagct cgcttggact cctgttgata gatccagtaa
     2581 tgacctaaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     2641 ttggtgagaa tccaagcctc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct
     2701 tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag
     2761 gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga
     2821 tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct
     2881 actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta
     2941 tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag
     3001 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
     3061 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
     3121 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
     3181 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     3241 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     3301 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     3361 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     3421 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     3481 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     3541 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     3601 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     3661 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     3721 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag
     3781 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     3841 tctcatgttt gacagcttat catcgataag cttccgatgg cg
//
