LOCUS       sequence-363359-        3446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L31p coding 
            region E8A 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: pRAR114
FEATURES             Location/Qualifiers
     source          1..3446
                     /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             1048..1077
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1093..1110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1166..1183)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1336..1382
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1478..1494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1478..1494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1491..1513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1502..1518
                     /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(1526..1532,1533..1550,1551..1556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1571..1592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1709..1726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1879..2467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1959..1978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2638..3429,3430..3446,1..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     3261..3280
                     /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 atgaagccca
      781 atatccatgc tgagtatcgt actgtggtgt tccacgacac cagtgttgat gagtacttta
      841 aaatcggctc gactatcaaa acagaccgtg agattgagct ggatggcgta acgtatccat
      901 acgtgacaat tgatgtctct tctaaatcgc acccgttcta tacagggaag ctgagaacag
      961 tggcatcaga aggaaatgtt gcacgattca cccaacgttt tggtcgtttt gttagcacga
     1021 aaaagggggc gtgagaagct tgggcccgaa caaaaactca tctcagaaga ggatctgaat
     1081 agcgccgtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
     1141 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1201 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1261 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     1321 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1381 atctgttgtt tgtcggtgaa ctctgaaagg aggaactata tccggatcga gatcctctag
     1441 agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1501 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1561 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1621 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1681 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1741 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1801 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaggccagga accgtaaaaa
     1861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2401 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2761 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3421 ttctgggtga gcaaaaacag gaaggc
//
