LOCUS       sequence-244671-        4399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative selection plasmid for evolution of an amber stop codon 
            suppression tRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4399)
  AUTHORS   Maranhao AC, Ellington AD
  TITLE     Evolving Orthogonal Suppressor tRNAs To Incorporate Modified Amino 
            Acids.
  JOURNAL   ACS Synth Biol. 2017 Jan 20;6(1):108-119. doi: 
            10.1021/acssynbio.6b00145. Epub 2016 Sep 21.
  PUBMED    27600875
REFERENCE   2  (bases 1 to 4399)
  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: 
            "10.1021/acssynbio.6b00145"; journalName: "ACS Synth Biol"; date: 
            "2017-01-20- 20"; volume: "6"; issue: "1"; pages: "108-119"
COMMENT     Sequence Label: pSELt_S7.A
FEATURES             Location/Qualifiers
     source          1..4399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(180..995)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(180..995)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     903..922
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     987..1006
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        complement(join(1657..1662,1663..1679,1680..1685))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      1797..2342
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2243..2262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2459..2476
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3025..3258
                     /codon_start=1
                     /gene="acpP"
                     /product="acyl carrier protein (ACP) from E. coli"
                     /label=ACP
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     CDS             4286..4315
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4331..4348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
       61 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
      121 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
      181 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
      241 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
      301 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
      361 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
      421 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
      481 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
      541 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
      601 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tctcacctga atcaggatat
      661 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
      721 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
      781 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
      841 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
      901 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
      961 ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     1021 taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1081 agcgtcagac cccgtagaaa cctcgacctg aatggaagcc ggcggcacct cgctaacgga
     1141 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc
     1201 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc
     1261 tcgggcagcg ttgggtcctg gccaatgcat accaaacctg cacgctagtt tcctgatgaa
     1321 catttttcca gcaattacac ctctgtcgat aattaactat tgacgaaaag ctgaaaacca
     1381 ctagaatgcg cctccgtggt accaattctt tttaagaatt gatggtatcg tggcgtagtt
     1441 cagcagggca gaacggcgat ctctaaaatc gcatgtcaga ggttcaaatc ctctcgtcgc
     1501 gaccaatttt gaaccccgct tcggcggggt tttttgtttt ctgtacattt cgtcaccctc
     1561 ccttcgcaat aaacgcccgt aatatgacca cgggtgcgca tgatcgtgca cacggtgcct
     1621 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     1681 gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     1741 gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga
     1801 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     1861 ggtttttcga aggttctctg aactaccaac tctttgaacc gaggtaactg gcttggagga
     1921 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     1981 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     2041 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     2101 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     2161 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     2221 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2281 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     2341 aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag
     2401 gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag
     2461 cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg
     2521 tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca
     2581 tagtaagcca gtatacactc cgctagcgct gatgtccggc ggtgcttttg ccgttacgca
     2641 ccaccccgtc agtagctgaa caggagggac agctgataga aacagaagcc actggagcac
     2701 ctcaaaaaca ccatcataca ctaaatcagt aagttggcag catcacccga cgcactttgc
     2761 gccgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct
     2821 gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta
     2881 gggccctcta gatgcatgct cgagcggcgg ccagtgtgat ggatatctgc agaattcgcc
     2941 cttcaaaaat cgatgagctg ttgacaatta atcatcgaac tagtttaatg tgtggaagcg
     3001 gccgctttca ggaggctcga gcatatgagc actatcgaag aacgcgttaa gaaaattatc
     3061 ggcgaacagc tgggcgttaa gcaggaagaa gttaccaaca atgcttcttt cgttgaagac
     3121 ctgggcgcgg attctcttga caccgttgag ctggtaatgg ctctggaaga agagtttgat
     3181 actgagattc cggacgaaga agctgagaaa atcaccaccg ttcaggctgc cattgattac
     3241 atcaacggcc accaggcgta gggcgcgccg ggcccgggct agtcacatct cgcagaactg
     3301 gttgccagtg cgaaggcggc cattagccag gcgtcagatg ttgccgcgtt agataatgtg
     3361 cgcgtcgaat atttgggtaa aaaagggcac ttaacccttc agatgacgac cctgcgtgag
     3421 ctgccgccag aagagcgtcc ggcagctggt gcggttatca acgaagcgaa agagcaggtt
     3481 cagcaggcgc tgaatgcgcg taaagcggaa ctggaaagcg ctgcactgaa tgcgcgtctg
     3541 gcggcggaaa cgattgatgt ctctctgcca ggtcgtcgca ttgaaaacgg cggtctgcat
     3601 ccggttaccc gtaccatcga ccgtatcgaa agtttcttcg gtgagcttgg ctttaccgtg
     3661 gcaaccgggc cggaaatcga agacgattat cataacttcg atgctctgaa cattcctggt
     3721 caccacccgg cgcgcgctga ccacgacact ttctggtttg acactacccg cctgctgcgt
     3781 acccagacct ctggcgtaca gatccgcacc atgaaagccc agcagccacc gattcgtatc
     3841 atcgcgcctg gccgtgttta tcgtaacgac tacgaccaga ctcacacgcc gatgttccat
     3901 cagatggaag gtctgattgt tgataccaac atcagcttta ccaacctgaa aggcacgctg
     3961 cacgacttcc tgcgtaactt ctttgaggaa gatttgcaga ttcgcttccg tccttcctac
     4021 ttcccgttta ccgaaccttc tgcagaagtg gacgtcatgg gtaaaaacgg taaatggctg
     4081 gaagtgctgg gctgcgggat ggtgcatccg aacgtgttgc gtaacgttgg catcgacccg
     4141 gaagtttact ctggtttcgg cttcgggatg gggatggagc gtctgactat gttgcgttac
     4201 ggcgtcaccg acctgcgttc attcttcgaa aacgatctgc gtttcctcaa acagtttaaa
     4261 taatagatct tcgaagcttg ggcccgaaca aaaactcatc tcagaagagg atctgaatag
     4321 cgccgtcgac catcatcatc atcatcattg agtttaaacg acgtccagct tggctgtttt
     4381 ggcggatgag agaagattt
//
