LOCUS       sequence-366945-        3882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  400 nt l31 promoter region, l31 5'UTR A74 and A76 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: pRAR104
FEATURES             Location/Qualifiers
     source          1..3882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(639..641,642..1355)
                     /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             639..1352
                     /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(669..697)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1292..1312
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1375..1404
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1420..1437
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1493..1510)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1663..1709
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1862..2521
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1928..1947)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2542..2636
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3043..3060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3177..3722)
                     /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(3257..3276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cactttatgc ttccggctga attctaaaga tctcggcaag gcatgaaaca gcacatcgcc
       61 aatcggatga tgatagtaat ctgccgccca tagcagcaat cgccagacgg agtgagtaaa
      121 caccggctca ctatccagca cttcgactac cgcttttagc tcattgagcg gcagttcgct
      181 ggcatcgcta actgatacca caatcccgat gcgctcctgc tgtttgccaa acggcacgcg
      241 cacgcgacac ccagctttaa ccgtcatgcc ttctggcagc agatagtcaa aggtacgagg
      301 aagcggaacg ggcaaggcaa cgtgggcaac gggcatagca tcatcctgac ttgaaattcg
      361 gtgggttagt atacacattg ccgtagaaga gtgcggatca gtttgcatac gctggttaat
      421 ttctgtatga tttcgcgcct tcgtacgaaa tgatcgtatt gaagctatac ttttaacatc
      481 gcgtggtgtc tggcgttagg gctggaggcg acgcggcctt aaaccgaggt tttcccatga
      541 aaaaagatat tcacccgaaa tacgaagaaa ttactgctag ctgctcttgc ggtaacgtaa
      601 tgaaaatccg ctccaccgtt ggtcatggtg gtggtggtat gagcaaagga gaagaacttt
      661 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
      721 ctgtccgtgg agagggtgaa ggtgatgcta caaacggaaa actcaccctt aaatttattt
      781 gcactactgg aaaactacct gttccgtggc caacacttgt cactactctg acctatggtg
      841 ttcaatgctt ttcccgttat ccggatcaca tgaaacggca tgactttttc aagagtgcca
      901 tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg acctacaaga
      961 cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag ttaaagggta
     1021 ttgattttaa agaagatgga aacattcttg gacacaaact cgagtacaac tttaactcac
     1081 acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaattc
     1141 gccacaacgt tgaagatggt tccgttcaac tagcagacca ttatcaacaa aatactccaa
     1201 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa tctgtccttt
     1261 cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta actgctgctg
     1321 ggattacaca tggcatggat gagctctaca aataaggatc tgaagcttgg gcccgaacaa
     1381 aaactcatct cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga
     1441 gtttaaacgg tctccagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
     1501 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
     1561 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     1621 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     1681 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaactg gatccttact
     1741 cgagtctaga ctgcagttga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
     1801 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     1861 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1921 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1981 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2041 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
     2101 tgagctggtg atatgggata gtgttaaccc ttgttacacc gttttccatg agcaaactga
     2161 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2221 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2281 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2341 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2401 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2461 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2521 atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc taagaactcc
     2581 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
     2641 gcctccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag
     2701 ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc
     2761 aaagtgcgtc gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt
     2821 gctccagtgg cttctgtttc tatcagctgt ccctcctgtt cagctactga cggggtggtg
     2881 cgtaacggca aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg
     2941 ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc
     3001 ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct
     3061 acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct
     3121 ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc
     3181 cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga
     3241 aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct
     3301 cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat
     3361 tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga
     3421 accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3481 ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag
     3541 tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc
     3601 ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc
     3661 cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc
     3721 aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt
     3781 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgtttgacag
     3841 cttatcatcg ataagcttcc gatggcgcgc cgagaggctt ta
//
