LOCUS       sequence-373311-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, scrambled 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: pRAR064
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(638..640,641..1354)
                     /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             638..1351
                     /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(668..696)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1291..1311
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1374..1403
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1419..1436
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1492..1509)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1662..1708
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1861..2520
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1927..1946)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2541..2635
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3042..3059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3176..3721)
                     /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(3256..3275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttatgcttc cggctgaatt ctaaagatct cggcaaggca tgaaacagca catcgccaat
       61 cggatgatga tagtaatctg ccgcccatag cagcaatcgc cagacggagt gagtaaacac
      121 cggctcacta tccagcactt cgactaccgc ttttagctca ttgagcggca gttcgctggc
      181 atcgctaact gataccacaa tcccgatgcg ctcctgctgt ttgccaaacg gcacgcgcac
      241 gcgacaccca gctttaaccg tcatgccttc tggcagcaga tagtcaaagg tacgaggaag
      301 cggaacgggc aaggcaacgt gggcaacggg catagcatca tcctgacttg aaattcggtg
      361 ggttagtata cacattgccg tagaagagtg cggatcagtt tgcatacgct ggttaatttc
      421 tgtatgattt ggcaggtttg acgggagata gggcacgcta gtcccggtgt tgtctcatct
      481 gtctcttacg gtaactgtag atcgtcccat catcaactcc taggaggaag gatctatgaa
      541 aaaagatatt cacccgaaat acgaagaaat tactgctagc tgctcttgcg gtaacgtaat
      601 gaaaatccgc tccaccgttg gtcatggtgg tggtggtatg agcaaaggag aagaactttt
      661 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
      721 tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa ctcaccctta aatttatttg
      781 cactactgga aaactacctg ttccgtggcc aacacttgtc actactctga cctatggtgt
      841 tcaatgcttt tcccgttatc cggatcacat gaaacggcat gactttttca agagtgccat
      901 gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga cctacaagac
      961 gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaagggtat
     1021 tgattttaaa gaagatggaa acattcttgg acacaaactc gagtacaact ttaactcaca
     1081 caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg
     1141 ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaacaaa atactccaat
     1201 tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat ctgtcctttc
     1261 gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg
     1321 gattacacat ggcatggatg agctctacaa ataaggatct gaagcttggg cccgaacaaa
     1381 aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag
     1441 tttaaacggt ctccagcttg gctgttttgg cggatgagag aagattttca gcctgataca
     1501 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
     1561 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     1621 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     1681 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaactgg atccttactc
     1741 gagtctagac tgcagttgat cgggcacgta agaggttcca actttcacca taatgaaata
     1801 agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa
     1861 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1921 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1981 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     2041 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
     2101 gagctggtga tatgggatag tgttaaccct tgttacaccg ttttccatga gcaaactgaa
     2161 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     2221 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     2281 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     2341 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     2401 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     2461 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     2521 tttgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct aagaactcca
     2581 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     2641 cctccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     2701 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     2761 aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
     2821 ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc
     2881 gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt
     2941 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     3001 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     3061 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     3121 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     3181 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     3241 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     3301 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     3361 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     3421 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3481 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     3541 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     3601 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     3661 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     3721 agaagatcat cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc
     3781 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gtttgacagc
     3841 ttatcatcga taagcttccg atggcgcgcc gagaggcttt acac
//
