LOCUS       sequence-369696-        3519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108 promoter, 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 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: pRAR074-control-sfGFP
FEATURES             Location/Qualifiers
     source          1..3519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..82
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     CDS             join(290..292,293..1006)
                     /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             290..1003
                     /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(320..348)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     943..963
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1026..1055
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1071..1088
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1144..1161)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1314..1360
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1513..2172
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVNPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFISFDLNVANM
                     DNFFAPVFTMGKYYTQGNKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1579..1598)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2193..2287
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2694..2711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2828..3373)
                     /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(2908..2927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccgagaggc tttacacttt atgcttccgg ctgaattcta aagatctctg acagctagct
       61 cagtcctagg tataatgcta gcggcaggtt tgacgggaga tagggcacgc tagtcccggt
      121 gttgtctcat ctgtctctta cggtaactgt agatcgtccc atcatcaact cctaggagga
      181 aggatctatg aaaaaagata ttcacccgaa atacgaagaa attactgcta gctgctcttg
      241 cggtaacgta atgaaaatcc gctccaccgt tggtcatggt ggtggtggta tgagcaaagg
      301 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
      361 gcacaaattt tctgtccgtg gagagggtga aggtgatgct acaaacggaa aactcaccct
      421 taaatttatt tgcactactg gaaaactacc tgttccgtgg ccaacacttg tcactactct
      481 gacctatggt gttcaatgct tttcccgtta tccggatcac atgaaacggc atgacttttt
      541 caagagtgcc atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg
      601 gacctacaag acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga
      661 gttaaagggt attgatttta aagaagatgg aaacattctt ggacacaaac tcgagtacaa
      721 ctttaactca cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa
      781 cttcaaaatt cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc attatcaaca
      841 aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca
      901 atctgtcctt tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt
      961 aactgctgct gggattacac atggcatgga tgagctctac aaataaggat ctgaagcttg
     1021 ggcccgaaca aaaactcatc tcagaagagg atctgaatag cgccgtcgac catcatcatc
     1081 atcatcattg agtttaaacg gtctccagct tggctgtttt ggcggatgag agaagatttt
     1141 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     1201 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     1261 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     1321 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaact
     1381 ggatccttac tcgagtctag actgcagttg atcgggcacg taagaggttc caactttcac
     1441 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     1501 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     1561 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     1621 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     1681 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca
     1741 atgaaagacg gtgagctggt gatatgggat agtgttaacc cttgttacac cgttttccat
     1801 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     1861 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     1921 gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt catcagtttt
     1981 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     2041 tatacgcaag gcaacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     2101 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     2161 ggcggggcgt aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac
     2221 ctaagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
     2281 tgagaatcca agcctccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
     2341 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
     2401 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
     2461 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
     2521 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
     2581 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
     2641 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
     2701 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     2761 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     2821 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     2881 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     2941 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     3001 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     3061 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     3121 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     3181 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     3241 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     3301 agaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3361 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     3421 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     3481 atgtttgaca gcttatcatc gataagcttc cgatggcgc
//
