LOCUS       sequence-363331-        3882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR G60 and C69 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: pRAR109
FEATURES             Location/Qualifiers
     source          1..3882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      156..701
                     /direction=RIGHT
                     /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     602..621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     818..835
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1242..1336)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1357..2016)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1931..1950
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(2169..2215)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2368..2385
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2441..2458)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2474..2503)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(2523..3236,3237..3239))
                     /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             complement(2526..3239)
                     /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(2566..2586)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3181..3209
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 cctctcggcg cgccatcgga agcttatcga tgataagctg tcaaacatga gaattacaac
       61 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
      121 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
      181 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
      241 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
      301 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
      361 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
      421 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
      481 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
      541 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
      601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
      661 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
      721 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
      781 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
      841 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
      901 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
      961 gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca gtagctgaac
     1021 aggagggaca gctgatagaa acagaagcca ctggagcacc tcaaaaacac catcatacac
     1081 taaatcagta agttggcagc atcacccgac gcactttgcg ccgaataaat acctgtgacg
     1141 gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg
     1201 gccaactttt ggcgaaaatg agacgttgat cggaggcttg gattctcacc aataaaaaac
     1261 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttcttaggt cattactgga
     1321 tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc
     1381 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     1441 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     1501 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     1561 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     1621 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     1681 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     1741 gtaacaaggg ttaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa
     1801 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     1861 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     1921 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     1981 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     2041 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     2101 acctcttacg tgcccgatca actgcagtct agactcgagt aaggatccag ttcaccgaca
     2161 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc
     2221 tggcagttcc ctactctcgc atggggagac cccacactac catcggcgct acggcgtttc
     2281 acttctgagt tcggcatggg gtcaggtggg accaccgcgc tactgccgcc aggcaaattc
     2341 tgttttatca gaccgcttct gcgttctgat ttaatctgta tcaggctgaa aatcttctct
     2401 catccgccaa aacagccaag ctggagaccg tttaaactca atgatgatga tgatgatggt
     2461 cgacggcgct attcagatcc tcttctgaga tgagtttttg ttcgggccca agcttcagat
     2521 ccttatttgt agagctcatc catgccatgt gtaatcccag cagcagttac aaactcaaga
     2581 aggaccatgt ggtcacgctt ttcgttggga tctttcgaaa ggacagattg tgtcgacagg
     2641 taatggttgt ctggtaaaag gacagggcca tcgccaattg gagtattttg ttgataatgg
     2701 tctgctagtt gaacggaacc atcttcaacg ttgtggcgaa ttttgaagtt agctttgatt
     2761 ccattctttt gtttgtctgc cgtgatgtat acattgtgtg agttaaagtt gtactcgagt
     2821 ttgtgtccaa gaatgtttcc atcttcttta aaatcaatac cctttaactc gatacgatta
     2881 acaagggtat caccttcaaa cttgacttca gcacgcgtct tgtaggtccc gtcatctttg
     2941 aaagatatag tgcgttcctg tacataacct tcgggcatgg cactcttgaa aaagtcatgc
     3001 cgtttcatgt gatccggata acgggaaaag cattgaacac cataggtcag agtagtgaca
     3061 agtgttggcc acggaacagg tagttttcca gtagtgcaaa taaatttaag ggtgagtttt
     3121 ccgtttgtag catcaccttc accctctcca cggacagaaa atttgtgccc attaacatca
     3181 ccatctaatt caacaagaat tgggacaact ccagtgaaaa gttcttctcc tttgctcata
     3241 ccaccaccac catgaccaac ggtggagcgg attttcatta cgttaccgca agagcagcta
     3301 gcagtaattt cttcgtattt cgggtgaata tcttttttca tgggaaaacc tcggtttaag
     3361 gccgcgtcgc tcttccaccc taacgcagac accacgcgat gttaaaagta tagcttcaat
     3421 acgatcattt cgtacgaagg cgcgaaatca tacagaaatt aaccagcgta tgcaaactga
     3481 tccgcactct tctacggcaa tgtgtatact aacccaccga atttcaagtc aggatgatgc
     3541 tatgcccgtt gcccacgttg ccttgcccgt tccgcttcct cgtacctttg actatctgct
     3601 gccagaaggc atgacggtta aagctgggtg tcgcgtgcgc gtgccgtttg gcaaacagca
     3661 ggagcgcatc gggattgtgg tatcagttag cgatgccagc gaactgccgc tcaatgagct
     3721 aaaagcggta gtcgaagtgc tggatagtga gccggtgttt actcactccg tctggcgatt
     3781 gctgctatgg gcggcagatt actatcatca tccgattggc gatgtgctgt ttcatgcctt
     3841 gccgagatct ttagaattca gccggaagca taaagtgtaa ag
//
