LOCUS       sequence-363358-        3446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L31p coding 
            region H6A 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: pRAR113
FEATURES             Location/Qualifiers
     source          1..3446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(56..160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     228..246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(284..306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     406..425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     619..641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     633..650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     634..650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        678..712
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     CDS             1051..1080
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1096..1113
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1169..1186)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1339..1385
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1481..1497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1481..1497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1494..1516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1505..1521
                     /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(1529..1535,1536..1553,1554..1559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1574..1595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1712..1729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1882..2470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1962..1981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2641..3432,3433..3446,1..55))
                     /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     3264..3283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
       61 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      121 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      181 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
      241 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      301 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      361 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      421 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      481 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      541 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      601 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt aagatctcgg
      661 ctgaattcta aagatctctg acagctagct cagtcctagg tataatgcta gcacgtcgac
      721 tctcgagtga gattgttgac ggtaccgtat tttggatcta ggaggaagga tctatgaagc
      781 ccaatatcgc gcctgagtat cgtactgtgg tgttccacga caccagtgtt gatgagtact
      841 ttaaaatcgg ctcgactatc aaaacagacc gtgagattga gctggatggc gtaacgtatc
      901 catacgtgac aattgatgtc tcttctaaat cgcacccgtt ctatacaggg aagctgagaa
      961 cagtggcatc agaaggaaat gttgcacgat tcacccaacg ttttggtcgt tttgttagca
     1021 cgaaaaaggg ggcgtgagaa gcttgggccc gaacaaaaac tcatctcaga agaggatctg
     1081 aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggtctc cagcttggct
     1141 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     1201 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     1261 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     1321 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1381 tttatctgtt gtttgtcggt gaactctgaa aggaggaact atatccggat cgagatcctc
     1441 tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1501 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1561 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1621 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1681 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1741 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1801 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaggcca ggaaccgtaa
     1861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3421 cgtttctggg tgagcaaaaa caggaa
//
