LOCUS       sequence-438920-        2694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amber Suppressor tRNA for Phenylalanine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2694)
  AUTHORS   O'Neil P
  TITLE     Amber Suppressor tRNA for Natural Amino Acids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 2694)
  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            "unpublished"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: pPhe57
FEATURES             Location/Qualifiers
     source          1..2694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..46)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     146..165
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(468..484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(468..484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(481..503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    492..508
                     /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(516..522,523..540,541..546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    561..582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(699..716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(870..1458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(950..969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1629..2420,2421..2489))
                     /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     2252..2271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2490..2594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2662..2680
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
       61 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      121 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
      181 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcat aggcgccgct
      241 tctttgagcg aacgatcaaa aataagtggg gccccatcaa aaaaatattc tcaacataaa
      301 aaactttgtg taatacttgt aacgctgaat tcgcccggat agctcagtcg gtagagcagg
      361 ggattctaaa tccccgtgtc cttggttcga ttccgagtcc gggcactgca gtctgcagat
      421 ccttagcgaa agctaaggat tttttttaag cttcgggcgt aatcatggtc atagctgttt
      481 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      541 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      601 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      661 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1741 tctggcccca gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca
     1801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2161 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2221 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2521 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2581 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     2641 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcg
//
