LOCUS       sequence-363333-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR U58C and U70C, 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: pRAR116
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(640..642,643..1356)
                     /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             640..1353
                     /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(670..698)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1293..1313
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1376..1405
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1421..1438
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1494..1511)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1664..1710
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1863..2522
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1929..1948)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2543..2637
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3044..3061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3178..3723)
                     /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(3258..3277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actttatgct tccggctgaa ttctaaagat ctcggcaagg catgaaacag cacatcgcca
       61 atcggatgat gatagtaatc tgccgcccat agcagcaatc gccagacgga gtgagtaaac
      121 accggctcac tatccagcac ttcgactacc gcttttagct cattgagcgg cagttcgctg
      181 gcatcgctaa ctgataccac aatcccgatg cgctcctgct gtttgccaaa cggcacgcgc
      241 acgcgacacc cagctttaac cgtcatgcct tctggcagca gatagtcaaa ggtacgagga
      301 agcggaacgg gcaaggcaac gtgggcaacg ggcatagcat catcctgact tgaaattcgg
      361 tgggttagta tacacattgc cgtagaagag tgcggatcag tttgcatacg ctggttaatt
      421 tctgtatgat ttcgcgcctt cgtacgaaat gatcgtattg aagctatact tttaacatcg
      481 cgtggtgtcc ggcgttaggg ccggaagagc gacgcggcct taaaccgagg ttttcccatg
      541 aaaaaagata ttcacccgaa atacgaagaa attactgcta gctgctcttg cggtaacgta
      601 atgaaaatcc gctccaccgt tggtcatggt ggtggtggta tgagcaaagg agaagaactt
      661 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      721 tctgtccgtg gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt
      781 tgcactactg gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt
      841 gttcaatgct tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc
      901 atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag
      961 acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt
     1021 attgatttta aagaagatgg aaacattctt ggacacaaac tcgagtacaa ctttaactca
     1081 cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     1141 cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca
     1201 attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt
     1261 tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct
     1321 gggattacac atggcatgga tgagctctac aaataaggat ctgaagcttg ggcccgaaca
     1381 aaaactcatc tcagaagagg atctgaatag cgccgtcgac catcatcatc atcatcattg
     1441 agtttaaacg gtctccagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     1501 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     1561 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     1621 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     1681 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaact ggatccttac
     1741 tcgagtctag actgcagttg atcgggcacg taagaggttc caactttcac cataatgaaa
     1801 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     1861 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     1921 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1981 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     2041 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     2101 gtgagctggt gatatgggat agtgttaacc cttgttacac cgttttccat gagcaaactg
     2161 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     2221 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     2281 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     2341 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     2401 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     2461 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     2521 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctaagaactc
     2581 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
     2641 agcctccgat caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca
     2701 gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg
     2761 caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg
     2821 tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg acggggtggt
     2881 gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat
     2941 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     3001 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     3061 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     3121 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     3181 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     3241 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     3301 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     3361 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     3421 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     3541 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     3601 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     3661 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     3721 caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct
     3781 tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc atgtttgaca
     3841 gcttatcatc gataagcttc cgatggcgcg ccgagaggct ttac
//
