LOCUS       sequence-366951-        3323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L34 coding 
            region, trrnB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3323)
  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 3323)
  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: pRAR119
FEATURES             Location/Qualifiers
     source          1..3323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(163..182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      976..1564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1465..1484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1717..1734
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1851..1872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1887..1892,1893..1910,1911..1917)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1925..1941
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1930..1952
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1949..1965
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1949..1965
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(2061..2107)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2260..2277
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2333..2350)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2366..2395)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2611..2645)
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     primer_bind     complement(2673..2690)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2673..2689)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2682..2704)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2898..2917)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3017..3039
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3077..3095)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3163..3267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3268..3323,1..13,14..805)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
       61 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      121 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      181 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      241 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      301 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      361 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      421 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      481 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      541 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      601 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      661 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      721 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      781 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
      841 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      901 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      961 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1021 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1081 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1141 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1201 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1261 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1321 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1381 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1441 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1501 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1561 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc tttgctggcc ttttgctcac
     1621 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     1681 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     1741 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     1801 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     1861 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     1921 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     1981 cttgcatgcc tgcaggtcga ctctagagga tctcgatccg gatatagttc ctcctttcag
     2041 agttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     2101 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg
     2161 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
     2221 ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg
     2281 aaaatcttct ctcatccgcc aaaacagcca agctggagac cgtttaaact caatgatgat
     2341 gatgatgatg gtcgacggcg ctattcagat cctcttctga gatgagtttt tgttcgggcc
     2401 caagcttctt acttagaaac ggtcagacga gcgcggcctt tagcacgacg acgtgccaga
     2461 acctgacgac catttttagt agccatacga gcacggaagc cgtgagaacg gttgcgcttc
     2521 agtacagacg gttgaaaagt gcgtttcata gatccttcct cctagatcca aaatacggta
     2581 ccgtcaacaa tctcactcga gagtcgacgt gctagcatta tacctaggac tgagctagct
     2641 gtcagagatc tttagaattc agccgagatc ttactggccg tcgttttaca acgtcgtgac
     2701 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2761 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2821 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2881 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     2941 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3001 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3061 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3121 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3181 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3241 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3301 cccttttttg cggcattttg cct
//
