LOCUS       sequence-421059-        2691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amber Suppressor tRNA for Lysine (Aspartate-derived).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2691)
  AUTHORS   O'Neil P
  TITLE     Amber Suppressor tRNA for Natural Amino Acids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 2691)
  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            "unpublished"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: pLys57-4
FEATURES             Location/Qualifiers
     source          1..2691
                     /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"
     primer_bind     complement(470..486)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(470..486)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(483..505)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    494..510
                     /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(518..524,525..542,543..548))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    563..584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(701..718)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(872..1460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(952..971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1631..2422,2423..2491))
                     /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     2254..2273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2492..2596)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2664..2682
                     /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 tgaattcgga gcggtagttc agtcggttag
      361 aataccagac tctaaatctg ggggtcgcgg gttcgagtcc cgtccgttcc gccactgcag
      421 atccttagcg aaagctaagg atttttttta agcttcgggc gtaatcatgg tcatagctgt
      481 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
      541 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      601 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      661 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      721 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      781 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      841 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      901 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      961 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1021 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1081 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1141 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1201 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1261 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1321 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1381 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1441 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1501 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1561 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1621 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1681 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1741 catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat
     1801 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1861 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1921 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1981 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2041 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2101 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2161 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2221 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2281 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2341 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2401 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2461 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2521 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2581 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     2641 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
