LOCUS       sequence-119488-        8213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phosphoserine orthogonal translation system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8213)
  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 8213)
  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: SepOTS-kappa
FEATURES             Location/Qualifiers
     source          1..8213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1298..1315)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1468..1554
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1646..1673
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             1869..2060
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2054..2076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2162..2304
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2217..2236
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2319..2336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2490..3078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2570..2589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3189..3208)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3200..4015
                     /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
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMSKLQFHLMLDEFF"
     primer_bind     complement(3273..3292)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        5115..5192
                     /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             5193..6275
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5212..5231)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6507..6512,6513..6529,6530..6536)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6544..6560
                     /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     6549..6571
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 taaagtatat tagttaagta taagaaggag atatacatat gggatccatg tctaaagaaa
       61 agtttgaacg tacaaaaccg cacgttaacg tcggtactat cggccacgtt gaccatggta
      121 aaacaacgct gaccgctgca atcactaccg tactggctaa aacctacggc ggtgctgctc
      181 gcgcattcga ccagatcgat aacgcgccgg aagaaaaagc tcgtggtatc accatcaaca
      241 cttctcgggt tgaatacgac accccgaccc gtcactacgc acacgtagac tgcccggggc
      301 acgccgacta tgttaaaaac atgatcaccg gtgctgcgca gatggacggc gcgatcctgg
      361 tagttgctgc gactgacggc ccgatgccgc agactcgtga gcacatcctg ctgggtcgtc
      421 aggtaggcgt tccgtacatc atcgtgttcc tgaacaaatg cgacatggtt gatgacgaag
      481 agctgctgga actggttgaa atggaagttc gtgaacttct gtctcagtac gacttcccgg
      541 gcgacgacac tccgatcgtt cgtggttctg ctctgaaagc gctggaaggc gacgcagagt
      601 gggaagcgaa aatcctggaa ctggctggct tcctggattc ttacattccg gaaccagagc
      661 gtgcgattga caagccgttc ctgctgccga tcaacggggt atactccatc tccggtcgtg
      721 gtaccgttgt ttcgggtcgt gtagaacgcg gtatcatcaa agttggtgaa gaagttgaaa
      781 tcgttggtat caaagagact cagaagtcta cctgtactgg cgttgaaatg ttccgcaaac
      841 tgctggacga aggccgtgct ggtgagtggg taggtgttct gctgcgtggt atcaaacgtg
      901 aagaaatcga acgtggtcag gtactggcta agccgggcac catcaagccg cacaccaagt
      961 tcgaatctga agtgtacatt ctgtccaaag atgaaggcgg ccgtcatact ccgttcttca
     1021 aaggctaccg tccgcagttc tacttccgta ctactgacgt gactggtacc atcgaactgc
     1081 cggaaggcgt agagatggta atgccgggcg acaacatcaa aatggttgtt accctgatcc
     1141 acccgatcgc gatggacgac ggtctgcgtt tcgcaatccg tgaaggcggc cgtaccgttg
     1201 gcgcgggcgt tgtagcaaaa gttctgagct aatggcgcgc cgtcgacctg caggcatgca
     1261 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
     1321 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
     1381 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     1441 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1501 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     1561 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     1621 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
     1681 ctacaaactc tttttgtttg ctagcttttc tctggtcccg ccgcatccat accgccagtt
     1741 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
     1801 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
     1861 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
     1921 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
     1981 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
     2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2161 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     2221 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc
     2281 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2401 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2461 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2521 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2581 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2641 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2701 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2761 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2821 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2881 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2941 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3001 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3061 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3121 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
     3181 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
     3241 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     3301 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     3361 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3421 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3481 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     3541 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3601 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3661 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3721 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3781 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3841 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3901 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3961 taatcctgat atgagtaaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
     4021 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4081 gcacatttcc ccgaaaagtg ccaggccgcg cttctttgag cgaacgatca aaaataagtg
     4141 gcgccccatc aaaaaaatat tctcaacata aaaaactttg tgtaatactt gtaacgctga
     4201 attcgccggg gtagtctagg ggttaggcag cggactctaa atccgcctta cgtgggttca
     4261 aatcccaccc ccggctccag gatcctctag agtcgacctg cagatcctta gcgaaagcta
     4321 aggatttttt ttgccgcggc gcttctttga gcgaacgatc aaaaataagt ggcgccccat
     4381 caaaaaaata ttctcaacat aaaaaacttt gtgtaatact tgtaacgctg aattcgccgg
     4441 ggtagtctag gggttaggca gcggactcta aatccgcctt acgtgggttc aaatcccacc
     4501 cccggctcca ggatcctcta gagtcgacct gcagatcctt agcgaaagct aaggattttt
     4561 tttgcggccc ggccgcgctt ctttgagcga acgatcaaaa ataagtggcg ccccatcaaa
     4621 aaaatattct caacataaaa aactttgtgt aatacttgta acgctgaatt cgccggggta
     4681 gtctaggggt taggcagcgg actctaaatc cgccttacgt gggttcaaat cccacccccg
     4741 gctccaggat cctctagagt cgacctgcag atccttagcg aaagctaagg attttttttg
     4801 ccgcggcgct tctttgagcg aacgatcaaa aataagtggc gccccatcaa aaaaatattc
     4861 tcaacataaa aaactttgtg taatacttgt aacgctgaat tcgccggggt agtctagggg
     4921 ttaggcagcg gactctaaat ccgccttacg tgggttcaaa tcccaccccc ggctccagga
     4981 tcctctagag tcgacctgca gatccttagc gaaagctaag gatttttttt gcggccgctt
     5041 tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg
     5101 catgcattta cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
     5161 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     5221 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     5281 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     5341 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     5401 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     5461 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     5521 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     5581 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     5641 actctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     5701 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     5761 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     5821 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     5881 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     5941 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     6001 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     6061 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     6121 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     6181 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6301 cgcgaattga tctggtttga cagcttatca tcgactgcac ggtgcaccaa tgcttctggc
     6361 gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc
     6421 gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt
     6481 tctggcaaat attctgaaat gagctgttga caattaatca tccggctcgt ataatgtgtg
     6541 gaattgtgag cggataacaa tttcacacag gaaacagacc catggcaatg tttaaaagag
     6601 aagaaatcat tgaaatggcc aataaggact ttgaaaaagc atggatcgaa actaaagacc
     6661 ttataaaagc taaaaagata aacgaaagtt acccaagaat aaaaccagtt tttggaaaaa
     6721 cacaccctgt aaatgacact attgaaaatt taagacaggc atatcttaga atgggttttg
     6781 aagaatatat aaacccagta attgtcgatg aaagagatat ttataaacaa ttcggcccag
     6841 aagctatggc agttttggat agatgctttt atttagcggg acttccaaga cctgacgttg
     6901 gtttgagcga tgaaaaaatt tcacagattg aaaaacttgg aattaaagtt tctgagcaca
     6961 aagaaagttt acaaaaaata cttcacggat acaaaaaagg aactcttgat ggtgacgatt
     7021 tagttttaga aatttcaaat gcacttgaaa tttcaagcga gatgggttta aaaattttag
     7081 aagatgtttt cccagaattt aaggatttaa ccgcagtttc ttcaaaatta actttaagaa
     7141 gccacatgac ttcaggatgg ttccttactg tttcagacct catgaacaaa aaacccttgc
     7201 catttaaact cttttcaatc gatagatgtt ttagaagaga acaaaaagaa gataaaagcc
     7261 acttaatgac ataccactct gcatcctgtg caattgcagg tgaaggcgtg gatattaatg
     7321 atggaaaagc aattgcagaa ggattattat cccaatttgg ctttacaaac tttaaattca
     7381 ttcctgatga aaagaaaagt aaatactaca cccctgaaac acagactgaa gtttacgcat
     7441 accacccaaa attaaaagaa tggctcgaag ttgctacatt tggagtatat tcgccagttg
     7501 cattaagcaa atacggaata gatgtacctg taatgaattt gggtcttggt gttgaaagac
     7561 ttgcaatgat ttctggaaat ttcgcagatg ttcgagaaat ggtatatcct cagttttacg
     7621 aacacaaact taatgaccgg aatgtcgctt caatggtaaa actcgataaa gttccagtaa
     7681 tggatgaaat ttacgattta acaaaagaat taattgagtc atgtgttaaa aacaaagatt
     7741 taaaatcccc ttgtgaatta gctattgaaa aaacgttttc atttggaaaa accaagaaaa
     7801 atgtaaaaat aaacattttt gaaaaagaag aaggtaaaaa tttactcgga ccttcaattt
     7861 taaacgaaat ctacgtttac gatggaaatg taattggaat tcctgaaagc tttgacggag
     7921 taaaagaaga atttaaagac ttcttagaaa aaggaaaatc agaaggggta gcaacaggca
     7981 ttcgatatat cgatgcgctt tgctttaaaa ttacttcaaa attagaagaa gcatttgtgt
     8041 caaacactac tgaattcaaa gttaaagttc caattgtcag aagtttaagc gacattaact
     8101 taaaaatcga tgatatcgca ttaaaacaga tcatgagcaa aaataaagta atcgacgtta
     8161 gaggcccagt ctttttaaat gtcgaagtaa aaattgaata agagctcggt acc
//
