LOCUS       sequence-363355-        3422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L31p coding 
            region with last 8 amino acids deleted, trrnB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3422)
  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 3422)
  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: pRAR111
FEATURES             Location/Qualifiers
     source          1..3422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(164..183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      977..1565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1466..1485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1718..1735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1852..1873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1888..1893,1894..1911,1912..1918)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1926..1942
                     /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     1931..1953
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1950..1966
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1950..1966
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(2062..2108)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2261..2278
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2334..2351)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2367..2396)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2711..2745)
                     /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(2773..2790)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2773..2789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2782..2804)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2998..3017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3117..3139
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3177..3195)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3263..3367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3368..3422,1..14,15..806)
                     /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 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
       61 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      121 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      181 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      241 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      301 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      361 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      421 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      481 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      541 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      601 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      661 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      721 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      781 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
      841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
      901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc ctttgctggc cttttgctca
     1621 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1681 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1741 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1801 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     1861 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1921 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1981 gcttgcatgc ctgcaggtcg actctagagg atctckrtcc ggatrtagtt cctcctttca
     2041 gagttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     2101 tttatttgat gcctggcagt tccctactct cgcatgggga gaccccacac taccatcggc
     2161 gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg cgctactgcc
     2221 gccaggcaaa ttctgtttta tcagaccgct tctgcgttct gatttaatct gtatcaggct
     2281 gaaaatcttc tctcatccgc caaaacagcc aagctggaga ccgtttaaac tcaatgatga
     2341 tgatgatgat ggtcgacggc gctattcaga tcctcttctg agatgagttt ttgttcgggc
     2401 ccaagcttct caacgaccaa aacgttgggt gaatcgtgca acatttcctt ctgatgccac
     2461 tgttctcagc ttccctgtat agaacgggtg cgatttagaa gagacatcaa ttgtcacgta
     2521 tggatacgtt acgccatcca gctcaatctc acggtctgtt ttgatagtcg agccgatttt
     2581 aaagtactca tcaacactgg tgtcgtggaa caccacagta cgatactcag gatggatatt
     2641 gggcttcata gatccttcct cctagatcca aaatacggta ccgtcaacaa tctcactcga
     2701 gagtcgacgt gctagcatta tacctaggac tgagctagct gtcagagatc tttagaattc
     2761 agccgagatc ttactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2821 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2881 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     2941 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     3001 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3061 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3121 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3181 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3241 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3301 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3361 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3421 cc
//
