LOCUS       sequence-421057-        2690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amber Suppressor tRNA for Lysine (Isoleucine-derived).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2690)
  AUTHORS   O'Neil P
  TITLE     Amber Suppressor tRNA for Natural Amino Acids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 2690)
  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"; journalName: 
            "unpublished"
COMMENT     Sequence Label: pLys57-3
FEATURES             Location/Qualifiers
     source          1..2690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     tRNA            338..413
                     /gene="ileX"
                     /product="E. coli tRNA-Ile recognizing AUA"
                     /label=ileX
     primer_bind     complement(469..485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(469..485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(482..504)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    493..509
                     /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(517..523,524..541,542..547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    562..583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(700..717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(871..1459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(951..970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1630..2421,2422..2490))
                     /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     2253..2272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2491..2595)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2663..2681
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcataggcg
      241 ccgcttcttt gagcgaacga tcaaaaataa gtggggcccc atcaaaaaaa tattctcaac
      301 ataaaaaact ttgtgtaata cttgtaacgc tgaattcggc cccttagctc agtggttaga
      361 gcaagcgact ctaaatcgct tggtcgctgg ttcaagtcca gcaggggcca ccactgcaga
      421 tccttagcga aagctaagga ttttttttaa gcttcgggcg taatcatggt catagctgtt
      481 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      541 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
      601 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1741 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     1801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     2641 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
