LOCUS       sequence-363342-        3299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L36 coding 
            region, trrnB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3299)
  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 3299)
  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: pRAR090-L36
FEATURES             Location/Qualifiers
     source          1..3299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(53..157)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     225..243
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(281..303)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     403..422
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     616..638
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     630..647
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     631..647
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        675..709
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     CDS             901..930
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             946..963
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1019..1036)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1189..1235
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1331..1347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1331..1347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1344..1366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1355..1371
                     /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(1379..1385,1386..1403,1404..1409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1424..1445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1562..1579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1732..2320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1812..1831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2491..3282,3283..3299,1..52))
                     /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     3114..3133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
       61 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      181 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
      241 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      301 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      361 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      421 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
      481 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
      541 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
      601 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtaag atctcggctg
      661 aattctaaag atctctgaca gctagctcag tcctaggtat aatgctagca cgtcgactct
      721 cgagtgagat tgttgacggt accgtatttt ggatctagga ggaaggatct atgaaagttc
      781 gtgcttccgt caagaaatta tgccgtaact gcaaaatcgt taagcgtgat ggtgtcatcc
      841 gtgtgatttg cagtgccgag ccgaagcata aacagcgcca aggctgagaa gcttgggccc
      901 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat
      961 cattgagttt aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc
     1021 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     1081 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     1141 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     1201 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaactctgaa
     1261 aggaggaact atatccggat cgagatcctc tagagtcgac ctgcaggcat gcaagcttgg
     1321 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1381 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1441 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1501 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1561 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcaactcact
     1621 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1681 caaaaggcca gcaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggc
//
