LOCUS       sequence-363334-        3519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108 promoter, 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 3519)
  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 3519)
  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: pRAR120
FEATURES             Location/Qualifiers
     source          1..3519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..75
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     CDS             join(283..285,286..999)
                     /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             283..996
                     /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(313..341)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     936..956
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1019..1048
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1064..1081
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1137..1154)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1307..1353
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1506..2165
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1572..1591)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2186..2280
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2687..2704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2821..3366)
                     /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(2901..2920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggctttacac tttatgcttc cggctgaatt ctaaagatct ctgacagcta gctcagtcct
       61 aggtataatg ctagccgcgc cttcgtacga aatgatcgta ttgaagctat acttttaaca
      121 tcgcgtggtg tctggcgtta gggctggaag agcgacgcgg ccttaaaccg aggttttccc
      181 atgaaaaaag atattcaccc gaaatacgaa gaaattactg ctagctgctc ttgcggtaac
      241 gtaatgaaaa tccgctccac cgttggtcat ggtggtggtg gtatgagcaa aggagaagaa
      301 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      361 ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt
      421 atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat
      481 ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt
      541 gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac
      601 aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag
      661 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac
      721 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa
      781 attcgccaca acgttgaaga tggttccgtt caactagcag accattatca acaaaatact
      841 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc
      901 ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct
      961 gctgggatta cacatggcat ggatgagctc tacaaataag gatctgaagc ttgggcccga
     1021 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca
     1081 ttgagtttaa acggtctcca gcttggctgt tttggcggat gagagaagat tttcagcctg
     1141 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     1201 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     1261 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     1321 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga actggatcct
     1381 tactcgagtc tagactgcag ttgatcgggc acgtaagagg ttccaacttt caccataatg
     1441 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1501 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1561 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1621 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1681 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     1741 acggtgagct ggtgatatgg gatagtgtta acccttgtta caccgttttc catgagcaaa
     1801 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1861 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1921 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1981 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     2041 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     2101 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2161 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctaagaa
     2221 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     2281 ccaagcctcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     2341 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     2401 gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
     2461 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
     2521 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     2581 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     2641 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     2701 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     2761 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     2821 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     2881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     2941 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     3001 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     3061 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3121 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     3181 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     3241 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     3301 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     3361 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     3421 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     3481 acagcttatc atcgataagc ttccgatggc gcgccgaga
//
