LOCUS       sequence-363356-        3446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L31p coding 
            region K2A mutation, trrnB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3446)
  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 3446)
  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: pRAR112
FEATURES             Location/Qualifiers
     source          1..3446
                     /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             1049..1078
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1094..1111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1167..1184)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1337..1383
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1479..1495)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1479..1495)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1492..1514)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1503..1519
                     /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(1527..1533,1534..1551,1552..1557))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1572..1593
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1710..1727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1880..2468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1960..1979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2639..3430,3431..3446,1..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     3262..3281
                     /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 tatggcgccc
      781 aatatccatc ctgagtatcg tactgtggtg ttccacgaca ccagtgttga tgagtacttt
      841 aaaatcggct cgactatcaa aacagaccgt gagattgagc tggatggcgt aacgtatcca
      901 tacgtgacaa ttgatgtctc ttctaaatcg cacccgttct atacagggaa gctgagaaca
      961 gtggcatcag aaggaaatgt tgcacgattc acccaacgtt ttggtcgttt tgttagcacg
     1021 aaaaaggggg cgtgagaagc ttgggcccga acaaaaactc atctcagaag aggatctgaa
     1081 tagcgccgtc gaccatcatc atcatcatca ttgagtttaa acggtctcca gcttggctgt
     1141 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1201 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1261 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1321 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1381 tatctgttgt ttgtcggtga actctgaaag gaggaactat atccggatcg agatcctcta
     1441 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     1501 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     1561 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     1621 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1681 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1741 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1801 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaggccagg aaccgtaaaa
     1861 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1921 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1981 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2041 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2101 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2221 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2281 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2341 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2401 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2461 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2521 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2581 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2641 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2701 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2761 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2821 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2881 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2941 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3001 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3061 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3121 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3181 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3241 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3301 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3361 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3421 tttctgggtg agcaaaaaca ggaagg
//
