LOCUS       sequence-363352-        3425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L31p coding 
            region with amino acids 2-8 deleted, trrnB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3425)
  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 3425)
  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: pRAR101
FEATURES             Location/Qualifiers
     source          1..3425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(54..158)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     226..244
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(282..304)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     404..423
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     617..639
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     631..648
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     632..648
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        676..710
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     CDS             1028..1057
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1073..1090
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1146..1163)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1316..1362
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1458..1474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1458..1474)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1471..1493)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1482..1498
                     /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)."
     promoter        complement(join(1506..1512,1513..1530,1531..1536))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1551..1572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1689..1706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1859..2447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1939..1958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2618..3409,3410..3425,1..53))
                     /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"
     primer_bind     3241..3260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
       61 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      121 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      181 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
      241 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
      301 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      361 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      421 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      481 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      541 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      601 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtaa gatctcggct
      661 gaattctaaa gatctctgac agctagctca gtcctaggta taatgctagc acgtcgactc
      721 tcgagtgaga ttgttgacgg taccgtattt tggatctagg aggaaggatc tatgtatcgt
      781 actgtggtgt tccacgacac cagtgttgat gagtacttta aaatcggctc gactatcaaa
      841 acagaccgtg agattgagct ggatggcgta acgtatccat acgtgacaat tgatgtctct
      901 tctaaatcgc acccgttcta tacagggaag ctgagaacag tggcatcaga aggaaatgtt
      961 gcacgattca cccaacgttt tggtcgtttt gttagcacga aaaagggggc gtgagaagct
     1021 tgggcccgaa caaaaactca tctcagaaga ggatctgaat agcgccgtcg accatcatca
     1081 tcatcatcat tgagtttaaa cggtctccag cttggctgtt ttggcggatg agagaagatt
     1141 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     1201 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     1261 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     1321 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1381 ctctgaaagg aggaactata tccggatcga gatcctctag agtcgacctg caggcatgca
     1441 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1501 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1561 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1621 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1681 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1741 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1801 atgtgagcaa aaggccagca aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3421 gaagg
//
