LOCUS       sequence-363325-        3872 bp    DNA     circular SYN 15-APR-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR with nt. 35-46 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 3872)
  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 3872)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pRAR040
FEATURES             Location/Qualifiers
     source          1..3872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(627..629,630..1343)
                     /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             627..1340
                     /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(657..685)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1280..1300
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1363..1392
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1408..1425
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1481..1498)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1651..1697
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1850..2509
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1916..1935)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2530..2624
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3031..3048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3165..3710)
                     /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(3245..3264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttatgctt ccggctgaat tctaaagatc tcggcaaggc atgaaacagc acatcgccaa
       61 tcggatgatg atagtaatct gccgcccata gcagcaatcg ccagacggag tgagtaaaca
      121 ccggctcact atccagcact tcgactaccg cttttagctc attgagcggc agttcgctgg
      181 catcgctaac tgataccaca atcccgatgc gctcctgctg tttgccaaac ggcacgcgca
      241 cgcgacaccc agctttaacc gtcatgcctt ctggcagcag atagtcaaag gtacgaggaa
      301 gcggaacggg caaggcaacg tgggcaacgg gcatagcatc atcctgactt gaaattcggt
      361 gggttagtat acacattgcc gtagaagagt gcggatcagt ttgcatacgc tggttaattt
      421 ctgtatgatt tcgcgccttc gtacgaaatg atcgtattga agctacgcgt ggtgtctggc
      481 gttagggctg gaagagcgac gcggccttaa accgaggttt tcccatgaaa aaagatattc
      541 acccgaaata cgaagaaatt actgctagct gctcttgcgg taacgtaatg aaaatccgct
      601 ccaccgttgg tcatggtggt ggtggtatga gcaaaggaga agaacttttc actggagttg
      661 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtccgtggag
      721 agggtgaagg tgatgctaca aacggaaaac tcacccttaa atttatttgc actactggaa
      781 aactacctgt tccgtggcca acacttgtca ctactctgac ctatggtgtt caatgctttt
      841 cccgttatcc ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt
      901 atgtacagga acgcactata tctttcaaag atgacgggac ctacaagacg cgtgctgaag
      961 tcaagtttga aggtgatacc cttgttaatc gtatcgagtt aaagggtatt gattttaaag
     1021 aagatggaaa cattcttgga cacaaactcg agtacaactt taactcacac aatgtataca
     1081 tcacggcaga caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacgttg
     1141 aagatggttc cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     1201 ctgtcctttt accagacaac cattacctgt cgacacaatc tgtcctttcg aaagatccca
     1261 acgaaaagcg tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg
     1321 gcatggatga gctctacaaa taaggatctg aagcttgggc ccgaacaaaa actcatctca
     1381 gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttaaacggtc
     1441 tccagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     1501 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     1561 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     1621 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     1681 tgggcctttc gttttatctg ttgtttgtcg gtgaactgga tccttactcg agtctagact
     1741 gcagttgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1801 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1861 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1921 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1981 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     2041 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     2101 atgggatagt gttaaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     2161 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     2221 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     2281 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     2341 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     2401 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2461 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga
     2521 gctcgcttgg actcctgttg atagatccag taatgaccta agaactccat ctggatttgt
     2581 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc ctccgatcaa
     2641 cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat
     2701 ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg
     2761 gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct
     2821 tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa
     2881 agcaccgccg gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg
     2941 agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca
     3001 gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     3061 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     3121 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     3181 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     3241 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     3301 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     3361 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     3421 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     3481 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     3541 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     3601 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     3661 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     3721 ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac
     3781 ctgaagtcag ccccatacga tataagttgt aattctcatg tttgacagct tatcatcgat
     3841 aagcttccga tggcgcgccg agaggcttta ca
//
