LOCUS       sequence-363324-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR, 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: pRAR017-L31-sfGFP
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(648..650,651..1364)
                     /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             648..1361
                     /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(678..706)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1301..1321
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1384..1413
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1429..1446
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1502..1519)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1672..1718
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1871..2530
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1937..1956)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2551..2645
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3052..3069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3186..3731)
                     /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(3266..3285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggctttacac tttatgcttc cggctgaatt ctaaagatct cggcaaggca tgaaacagca
       61 catcgccaat cggatgatga tagtaatctg ccgcccatag cagcaatcgc cagacggagt
      121 gagtaaacac cggctcacta tccagcactt cgactaccgc ttttagctca ttgagcggca
      181 gttcgctggc atcgctaact gataccacaa tcccgatgcg ctcctgctgt ttgccaaacg
      241 gcacgcgcac gcgacaccca gctttaaccg tcatgccttc tggcagcaga tagtcaaagg
      301 tacgaggaag cggaacgggc aaggcaacgt gggcaacggg catagcatca tcctgacttg
      361 aaattcggtg ggttagtata cacattgccg tagaagagtg cggatcagtt tgcatacgct
      421 ggttaatttc tgtatgattt cgcgccttcg tacgaaatga tcgtattgaa gctatacttt
      481 taacatcgcg tggtgtctgg cgttagggct ggaagagcga cgcggcctta aaccgaggtt
      541 ttcccatgaa aaaagatatt cacccgaaat acgaagaaat tactgctagc tgctcttgcg
      601 gtaacgtaat gaaaatccgc tccaccgttg gtcatggtgg tggtggtatg agcaaaggag
      661 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
      721 acaaattttc tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa ctcaccctta
      781 aatttatttg cactactgga aaactacctg ttccgtggcc aacacttgtc actactctga
      841 cctatggtgt tcaatgcttt tcccgttatc cggatcacat gaaacggcat gactttttca
      901 agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga
      961 cctacaagac gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt
     1021 taaagggtat tgattttaaa gaagatggaa acattcttgg acacaaactc gagtacaact
     1081 ttaactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact
     1141 tcaaaattcg ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaacaaa
     1201 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat
     1261 ctgtcctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa
     1321 ctgctgctgg gattacacat ggcatggatg agctctacaa ataaggatct gaagcttggg
     1381 cccgaacaaa aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat
     1441 catcattgag tttaaacggt ctccagcttg gctgttttgg cggatgagag aagattttca
     1501 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     1561 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     1621 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     1681 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaactgg
     1741 atccttactc gagtctagac tgcagttgat cgggcacgta agaggttcca actttcacca
     1801 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     1861 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     1921 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     1981 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     2041 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     2101 gaaagacggt gagctggtga tatgggatag tgttaaccct tgttacaccg ttttccatga
     2161 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     2221 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     2281 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     2341 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     2401 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     2461 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     2521 cggggcgtaa tttgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct
     2581 aagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     2641 agaatccaag cctccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
     2701 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
     2761 attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt
     2821 ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac
     2881 ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg
     2941 cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa
     3001 ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac
     3061 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg
     3121 agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag
     3181 ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag
     3241 tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc
     3301 gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt
     3361 ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg
     3421 tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga
     3481 gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta attgatttag
     3541 aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg
     3601 actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg
     3661 aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa
     3721 aacgatctca agaagatcat cttattaatc agataaaata tttctagatt tcagtgcaat
     3781 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     3841 gtttgacagc ttatcatcga taagcttccg atggcgcgcc gaga
//
