LOCUS       sequence-363339-        3323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L36p coding 
            region, trrnB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3323)
  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 3323)
  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: pRAR089-L36p
FEATURES             Location/Qualifiers
     source          1..3323
                     /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             928..957
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             973..990
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1046..1063)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1216..1262
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1358..1374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1358..1374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1371..1393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1382..1398
                     /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(1406..1412,1413..1430,1431..1436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1451..1472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1589..1606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1759..2347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1839..1858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2518..3309,3310..3323,1..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     3141..3160
                     /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 tctatgaaag
      781 ttcttaactc tctgcgtacc gcaaaagaac gccatccaga ctgtcagatt gtgaagcgaa
      841 aaggacggct atatgtgatt tgtaaatcta atccacgttt taaggccgtt cagggtcgta
      901 agaaaaaacg ttgagaagct tgggcccgaa caaaaactca tctcagaaga ggatctgaat
      961 agcgccgtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
     1021 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1081 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1141 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     1201 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1261 atctgttgtt tgtcggtgaa ctctgaaagg aggaactata tccggatcga gatcctctag
     1321 agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaggccagga accgtaaaaa
     1741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atcacagttc
     1981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2161 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2341 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2461 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2521 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2581 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2641 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2701 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2761 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2821 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2881 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2941 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3001 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3061 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3121 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3181 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3241 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3301 ttctgggtga gcaaaaacag gaa
//
