LOCUS       sequence-313679-        7530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding tRNA-SupD for serine amber suppression. *Update: 
            Please use the updated version, Plasmid 198598: SupD v2, which is 
            more stable..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7530)
  AUTHORS   Pirman NL, Barber KW, Aerni HR, Ma NJ, Haimovich AD, Rogulina S, 
            Isaacs FJ, Rinehart J
  TITLE     A flexible codon in genomically recoded Escherichia coli permits 
            programmable protein phosphorylation.
  JOURNAL   Nat Commun. 2015 Sep 9;6:8130. doi: 10.1038/ncomms9130.
  PUBMED    26350500
REFERENCE   2  (bases 1 to 7530)
  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: "Nat 
            Commun."; date: "2015-09-9"; volume: "6:8130. doi"; pages: " 
            10.1038/ncomms9130"
COMMENT     Sequence Label: SupD
FEATURES             Location/Qualifiers
     source          1..7530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      139..225
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      317..344
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             540..731
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(725..747)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    833..972
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     888..907
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(987..1004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1158..1746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1238..1257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1857..1876)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1868..2683
                     /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
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVAHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMSKLQFHLMLDEFF"
     primer_bind     complement(1941..1960)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3104..3181
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3182..4264
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDDSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLATIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3201..3220)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4496..4501,4502..4518,4519..4525)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4533..4549
                     /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)."
     primer_bind     4538..4560
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(7499..7516)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
       61 cgaactcaga agtgaaaggc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
      121 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
      181 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
      241 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
      301 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
      361 ctttttgttt gctagctttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
      421 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
      481 ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag
      541 tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa
      601 cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc
      661 ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg
      721 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
      781 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
      841 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
      901 gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt aaggagaaaa
      961 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1021 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1081 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1141 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1201 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1261 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1321 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1381 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1441 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1501 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1561 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     1621 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1681 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1741 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1801 cgttaaggga ttttggtcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1861 gggtgttatg agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa
     1921 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1981 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2041 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2101 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2161 cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2221 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2281 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2341 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2401 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtcg ctcatggtga
     2461 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2521 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2581 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2641 gagtaaattg cagtttcatt tgatgctcga tgagtttttc taagaattaa ttcatgagcg
     2701 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2761 gaaaagtgcc accgcggccg cgcttctttg agcgaacgat caaaaataag tggcgcccca
     2821 tcaaaaaaat attctcaaca taaaaaactt tgtgtaatac ttgtaacgct gaattcggag
     2881 agatgccgga gcggctgaac ggaccggtct ctaaaaccgg agtaggggca actctaccgg
     2941 gggttcaaat ccccctctct ccgccaggat cctctagagt cgacctgcag atccttagcg
     3001 aaagctaagg attttttttg cggccgcttt caccgtcatc accgaaacgc gcgaggcagc
     3061 agatcaattc gcgcgcgaag gcgaagcggc atgcatttac gttgacacca tcgaatggtg
     3121 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     3181 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     3241 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     3301 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     3361 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     3421 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     3481 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     3541 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     3601 cactaatgtt ccggcgttat ttcttgatga ctctgaccag acacccatca acagtattat
     3661 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     3721 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     3781 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     3841 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     3901 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     3961 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     4021 atgttatatc ccgccgttag ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     4081 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     4141 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     4201 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     4261 gtgagcgcaa cgcaattaat gtaagttagc gcgaattgat ctggtttgac agcttatcat
     4321 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     4381 tgtgcaggtc gtagatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     4441 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     4501 aattaatcat ccggcccgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
     4561 aaacagacca tggcaatgtt taaaagagaa gaaatcattg aaatggccaa taaggacttt
     4621 gaaaaagcat ggatcgaaac taaagacctt ataaaagcta aaaagataaa cgaaagttac
     4681 ccaagaataa aaccagtttt tggaaaaaca caccctgtaa atgacactat tgaaaattta
     4741 agacaggcat atcttagaat gggttttgaa gaatatataa acccagtaat tgtcgatgaa
     4801 agagatattt ataaacaatt cggcccagaa gctatggcag ttttggatag atgcttttat
     4861 ttagcgggac ttccaagacc tgacgttggt ttgagcgatg aaaaaatttc acagattgaa
     4921 aaacttggaa ttaaagtttc tgagcacaaa gaaagtttac aaaaaatact tcacggatac
     4981 aaaaaaggaa ctcttgatgg tgacgattta gttttagaaa tttcaaatgc acttgaaatt
     5041 tcaagcgaga tgggtttaaa aattttagaa gatgttttcc cagaatttaa ggatttaacc
     5101 gcagtttctt caaaattaac tttaagaagc cacatgactt caggatggtt ccttactgtt
     5161 tcagacctca tgaacaaaaa acccttgcca tttaaactct tttcaatcga tagatgtttt
     5221 agaagagaac aaaaagaaga taaaagccac ttaatgacat accactctgc atcctgtgca
     5281 attgcaggtg aaggcgtgga tattaatgat ggaaaagcaa ttgcagaagg attattatcc
     5341 caatttggct ttacaaactt taaattcatt cctgatgaaa agaaaagtaa atactacacc
     5401 cctgaaacac agactgaagt ttacgcatac cacccaaaat taaaagaatg gctcgaagtt
     5461 gctacatttg gagtatattc gccagttgca ttaagcaaat acggaataga tgtacctgta
     5521 atgaatttgg gtcttggtgt tgaaagactt gcaatgattt ctggaaattt cgcagatgtt
     5581 cgagaaatgg tatatcctca gttttacgaa cacaaactta atgaccggaa tgtcgcttca
     5641 atggtaaaac tcgataaagt tccagtaatg gatgaaattt acgatttaac aaaagaatta
     5701 attgagtcat gtgttaaaaa caaagattta aaatcccctt gtgaattagc tattgaaaaa
     5761 acgttttcat ttggaaaaac caagaaaaat gtaaaaataa acatttttga aaaagaagaa
     5821 ggtaaaaatt tactcggacc ttcaatttta aacgaaatct acgtttacga tggaaatgta
     5881 attggaattc ctgaaagctt tgacggagta aaagaagaat ttaaagactt cttagaaaaa
     5941 ggaaaatcag aaggggtagc aacaggcatt cgatatatcg atgcgctttg ctttaaaatt
     6001 acttcaaaat tagaagaagc atttgtgtca aacactactg aattcaaagt taaagttcca
     6061 attgtcagaa gtttaagcga cattaactta aaaatcgatg atatcgcatt aaaacagatc
     6121 atgagcaaaa ataaagtaat cgacgttaga ggcccagtct ttttaaatgt cgaagtaaaa
     6181 attgaataag agctcggtac ctaaagtata ttagttaagt ataagaagga gatatacata
     6241 tgggatccat gtctaaagaa aagtttgaac gtacaaaacc gcacgttaac gtcggtacta
     6301 tcggccacgt tgaccatggt aaaacaacgc tgaccgctgc aatcactacc gtactggcta
     6361 aaacctacgg cggtgctgct cgcgcattcg accagatcga taacgcgccg gaagaaaaag
     6421 ctcgtggtat caccatcaac acttctcggg ttgaatacga caccccgacc cgtcactacg
     6481 cacacgtaga ctgcccgggg cacgccgact atgttaaaaa catgatcacc ggtgctgcgc
     6541 agatggacgg cgcgatcctg gtagttgctg cgactgacgg cccgatgccg cagactcgtg
     6601 agcacatcct gctgggtcgt caggtaggcg ttccgtacat catcgtgttc ctgaacaaat
     6661 gcgacatggt tgatgacgaa gagctgctgg aactggttga aatggaagtt cgtgaacttc
     6721 tgtctcagta cgacttcccg ggcgacgaca ctccgatcgt tcgtggttct gctctgaaag
     6781 cgctggaagg cgacgcagag tgggaagcga aaatcctgga actggctggc ttcctggatt
     6841 cttacattcc ggaaccagag cgtgcgattg acaagccgtt cctgctgccg atcaacgggg
     6901 tatactccat ctccggtcgt ggtaccgttg tttcgggtcg tgtagaacgc ggtatcatca
     6961 aagttggtga agaagttgaa atcgttggta tcaaagagac tcagaagtct acctgtactg
     7021 gcgttgaaat gttccgcaaa ctgctggacg aaggccgtgc tggtgagtgg gtaggtgttc
     7081 tgctgcgtgg tatcaaacgt gaagaaatcg aacgtggtca ggtactggct aagccgggca
     7141 ccatcaagcc gcacaccaag ttcgaatctg aagtgtacat tctgtccaaa gatgaaggcg
     7201 gccgtcatac tccgttcttc aaaggctacc gtccgcagtt ctacttccgt actactgacg
     7261 tgactggtac catcgaactg ccggaaggcg tagagatggt aatgccgggc gacaacatca
     7321 aaatggttgt taccctgatc cacccgatcg cgatggacga cggtctgcgt ttcgcaatcc
     7381 gtgaaggcgg ccgtaccgtt ggcgcgggcg ttgtagcaaa agttctgagc taatggcgcg
     7441 ccgtcgacct gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc
     7501 tgatacagat taaatcagaa cgcagaagcg
//
