LOCUS       sequence-198038-        6434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tryptophanyl tRNA synthetase (ScWRS) and supressor 
            tryptophanyl tRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6434)
  AUTHORS   Abil Z, Ellefson JW, Gollihar JD, Watkins E, Ellington AD
  TITLE     Compartmentalized partnered replication for the directed evolution 
            of genetic parts and circuits.
  JOURNAL   Nat Protoc. 2017 Dec;12(12):2493-2512. doi: 10.1038/nprot.2017.119. 
            Epub 2017 Nov 9.
  PUBMED    29120463
REFERENCE   2  (bases 1 to 6434)
  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: 
            "10.1038/nprot.2017.119"; journalName: "Nat Protoc"; date: 
            "2017-12"; volume: "12"; issue: "12"; pages: "2493-2512"
COMMENT     Sequence Label: pRST11B-AS3_4
FEATURES             Location/Qualifiers
     source          1..6434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(126..714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(206..225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(885..1676,1677..1745))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1508..1527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1746..1850)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1884..1961
                     /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             1962..3044
                     /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
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1981..2000)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3236..3241,3242..3257,3258..3264)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3272..3288
                     /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)."
     CDS             5941..6132
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6126..6148)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6234..6374
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6289..6308
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6389..6406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
       61 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      541 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
      601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1201 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1501 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1861 cgaaaagtgc cacctgacgt cccgacacca tcgaatggtg caaaaccttt cgcggtatgg
     1921 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     1981 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     2041 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     2101 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     2161 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     2221 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     2281 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     2341 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     2401 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     2461 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     2521 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     2581 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     2641 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     2701 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     2761 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     2821 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     2881 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     2941 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3001 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3061 gtaagttagc tcactcatta ggcacaattc tcatgtttga cagcttatca tcgactgcac
     3121 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
     3181 cgtaaatcac tgcataattc gtgtcgctca aggcgcaaag cttgtcgacg agctgttgac
     3241 aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacaagg
     3301 agatctagaa tgagcaacga cgaaactgta gagaaagtca cccaacaagt gtcggaacta
     3361 aaaagcacag atgttaaaga gcaagtagtt acaccttggg atgtggaagg tggggttgat
     3421 gaacaaggta gagcccaaaa tattgattac gacaaattga tcaaacaatt cggtactaag
     3481 ccggtcaacg aagaaaccct gaagagattc aagcaagtga cgggtcgtga accacatcat
     3541 tttttgcgta agggattgtt tttcagtgag cgtgacttca ctaaaatatt agacctttac
     3601 gaacaaggca aaccattttt cctatacact ggtagaggtc cttcgagcga ttctatgcac
     3661 ttgggtcata tgatcccttt tgtcttcacc aaatggttac aggaagtgtt tgacgtacca
     3721 ttagtcatag agttgacaga tgacgaaaaa tttttattca aacacaagtt gaccatcaat
     3781 gacgttaaga attttgcccg tgaaaatgcc aaggatatca ttgctgttgg ctttgatcca
     3841 aagaacacct ttatcttttc tgatttgcaa tacatgggtg gtgcatttta cgaaactgta
     3901 gtaagagttt ccagacaaat tacaggatcc actgcaaagg ctgttttcgg gtttaatgac
     3961 tccgactgta ttggtaagtt ccattttgcc tccattcaaa ttgctaccgc attcccaagc
     4021 tcatttccta atgtgttagg cttgcctgat aagacaccat gtttgattcc atgtgcaatt
     4081 gaccaagatc catatttcag agtttgtagg gatgtcgcgg ataaattgaa gtactccaaa
     4141 cctgctttgc ttcattccag attctttcca gctttgcaag gttccacgac caaaatgtca
     4201 gcctctgatg ataccactgc cattttcatg accgatacac caaagcaaat tcaaaagaaa
     4261 attaacaagt acgcattcag cggtggtcaa gtgtccgccg acctacatag agaattaggt
     4321 ggtaatcccg atgtcgatgt tgcataccaa tacttgtcat ttttcaagga tgacgatgtt
     4381 ttcttgaagg aatgctatga caaatataag tccggtgaat tactatcagg tgaaatgaag
     4441 aaactgtgta tcgagactct gcaagaattc gttaaggcgt tccaggaacg cagagctcag
     4501 gtggacgaag agaccttgga caaattcatg gtcccacata agttggtttg gggcgaaaag
     4561 gaaagacttg tcgcacctaa gccaaaaact aagcaagaaa agaagtaact cgagagagaa
     4621 tataaaaagc cagattatta atccggcttt tttattattt cggccgacgc gctgggctac
     4681 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     4741 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     4801 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga
     4861 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
     4921 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     4981 cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa
     5041 gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga
     5101 acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg
     5161 ggtcctggcc aatgcatacc aaacctgcac gctagtttcc tgatgaacat ttttccagca
     5221 attacacctc tgtcgataat taactattga cgaaaagctg aaaaccacta gaatgcgcct
     5281 ccgtggtacc aattcttttt aagaattgat ggtatgaagc ggtggctcaa tggtagagct
     5341 ggcgcctcta aagcgccggg ttgcaggttc aattcctgtc cgtttcacca attttgaacc
     5401 ccgcttcggc ggggtttttt gttttctgta catttcgtca ccctcccttc gcaataaacg
     5461 cccgtaatat ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc
     5521 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
     5581 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
     5641 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc
     5701 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg
     5761 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc
     5821 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc
     5881 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca
     5941 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
     6001 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
     6061 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
     6121 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6181 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     6241 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     6301 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     6361 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6421 ggctgcggcg agcg
//
