LOCUS       sequence-363344-        3446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23108(spel), pHP14 stability hairpin, strong RBS, L31p coding 
            region, 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: pRAR091-L31p
FEATURES             Location/Qualifiers
     source          1..3446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(50..154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     222..240
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(278..300)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     400..419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     613..635
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     627..644
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     628..644
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        672..706
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     CDS             1045..1074
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1090..1107
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1163..1180)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1333..1379
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1475..1491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1475..1491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1488..1510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1499..1515
                     /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(1523..1529,1530..1547,1548..1553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1568..1589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1706..1723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1956..1975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2635..3426,3427..3446,1..49))
                     /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     3258..3277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
       61 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      121 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
      181 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      241 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      301 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      361 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      421 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      481 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      541 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      601 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtaagatc tcggctgaat
      661 tctaaagatc tctgacagct agctcagtcc taggtataat gctagcacgt cgactctcga
      721 gtgagattgt tgacggtacc gtattttgga tctaggagga aggatctatg aagcccaata
      781 tccatcctga gtatcgtact gtggtgttcc acgacaccag tgttgatgag tactttaaaa
      841 tcggctcgac tatcaaaaca gaccgtgaga ttgagctgga tggcgtaacg tatccatacg
      901 tgacaattga tgtctcttct aaatcgcacc cgttctatac agggaagctg agaacagtgg
      961 catcagaagg aaatgttgca cgattcaccc aacgttttgg tcgttttgtt agcacgaaaa
     1021 agggggcgtg agaagcttgg gcccgaacaa aaactcatct cagaagagga tctgaatagc
     1081 gccgtcgacc atcatcatca tcatcattga gtttaaacgg tctccagctt ggctgttttg
     1141 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     1201 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     1261 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     1321 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1381 tgttgtttgt cggtgaactc tgaaaggagg aactatatcc ggatcgagat cctctagagt
     1441 cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1501 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1561 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1621 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1681 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1741 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1801 ataacgcagg aaagaacatg tgagcaaaag gccagcaaag gccaggaacc gtaaaaaggc
     1861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2761 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3421 tgggtgagca aaaacaggaa ggcaaa
//
