LOCUS       sequence-104810-        8086 bp    DNA     circular SYN 14-APR-2025
DEFINITION  This plasmid encodes the mouse Mars gene with the L274G mutation 
            under a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8086)
  AUTHORS   Mahdavi A, Hamblin GD, Jindal GA, Bagert JD, Dong C, Sweredoski MJ, 
            Hess S, Schuman EM, Tirrell DA
  TITLE     Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of 
            Mammalian Protein Synthesis.
  JOURNAL   J Am Chem Soc. 2016 Apr 6;138(13):4278-81. doi: 
            10.1021/jacs.5b08980. Epub 2016 Mar 25.
  PUBMED    26991063
REFERENCE   2  (bases 1 to 8086)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/jacs.5b08980"; journalName: "J Am Chem Soc"; date: 
            "2016-04-6- 6"; volume: "138"; issue: "13"; pages: "4278-81"
COMMENT     Sequence Label: pMarsL274G
FEATURES             Location/Qualifiers
     source          1..8086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             913..930
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2995..3006
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(3247..3258)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3680..3697)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3686..3910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3956..4384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4043..4062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4253..4274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4393..4413)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4398..4727
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4578..4713
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4640..4659
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4794..5588
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4848..4867)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5458..5477
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5762..5883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5799..5818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5853..5872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5932..5948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5932..5948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5945..5967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5956..5972
                     /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(5980..5986,5987..6004,6005..6010))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6025..6046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6163..6180)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6334..6919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6414..6433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7090..7881,7882..7950))
                     /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     7713..7732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7951..8055)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg cccaccatca ccatcaccat atgagactgt tcgtgagcga gggttccccg
      961 gggagcctgc ccgtgctggc tgcggccgcg agggcccggg gtcgggcgga gctgctcatc
     1021 agcaccgtag gccccgaaga gtgtgtggta ccattcctta cccggcctaa ggtccctgtc
     1081 ttgcagctgg atagtggcaa ctacctcttc tctgctagtg caatctgccg atattttttt
     1141 ctgttatgtg gctgggaaca agatgatctc accaaccagt ggctggaatg ggaggcaaca
     1201 gaactgcagc cagttctgtc tgctgcccta cactgtctag tggttcaagg caagaaaggg
     1261 gaagatatac ttggcccact tcggagagtc ctgactcaca ttgatcacag cttgagtcgt
     1321 caaaactgtc ctttcctggc tggggacaca gaatctctag ctgacattgt tttgtgggga
     1381 gcactgtatc ctttactgca agacccagct tacctccctg aggagttggg tgccctgcaa
     1441 agttggttcc agacactgag tacccaggaa ccgtgtcagc gagctgcaga gacggtgcta
     1501 aaacagcagg gtgtcctggc acttcgtctg tacctccaga aacagccaca gcctcagccc
     1561 ccgcctcctg aggggagaac tgtcagcaac gagctggagg aagaggaact ggctaccttg
     1621 tctgaggagg acatcgttac agctgttgcc gcgtgggaga agggtctgga aagcctgcct
     1681 ccgctaaagc tccagcagca tccagtgttg cctgtgcctg gagagaggaa tgttctcatc
     1741 accagtgccg gaccctatgt caacaatgtc ccccaccttg gaaacatcat tggctgtgtg
     1801 ctcagtgctg atgtctttgc aaggtattgt cgccttcgcc agtggaatac cctctatctg
     1861 tgtggtacag atgagtatgg tactgcgaca gagaccaagg ccatggagga gggcctaacc
     1921 ccacgggaaa tctgtgacaa gtaccatgcc atccatgctg acatctaccg ctggttcggc
     1981 atatcgttcg atactttcgg gcgcactacc actcctcagc agaccaaaat cacccaggac
     2041 atcttccaga ggttgctgac ccgggggttt gtgctgcgag atactgtgga gcagcttcgg
     2101 tgtgagcggt gtgcacgttt cctggctgac cgctttgtgg agggtgtgtg tcccttctgt
     2161 ggctatgaag aggcccgagg tgaccagtgt gacaggtgtg gcaagctcat caatgccatt
     2221 gagctcaaga aaccacagtg caaaatctgc cgctcctgcc ctgtggtgag gtcctcacag
     2281 cacctgtttc tagacttgcc taagttggaa aagcgtctgg aggactggtt ggggaagaca
     2341 gtgcctggca gtgactggac acccaatgcc aggttcatta tacgttcctg gcttcgagat
     2401 ggcctcaagc cacgatgcat caccagagac ctcaaatggg gaacgcctgt gcccttggaa
     2461 ggttttgagg acaaggtatt ttacgtctgg tttgatgcta ctattggcta cgtgtccatc
     2521 acagccaact acacagacca atgggagaaa tggtggaaga acccagaaca agtggacctt
     2581 taccagttca tggccaaaga caatgttccc ttccatggct tggtctttcc gtgttcagtc
     2641 ctaggagctg aggacaacta caccctggtc aagcacatca ttgctacaga gtacctgaac
     2701 tatgaggatg ggaaattctc taagagccgg ggcataggag tgtttggaga catggccaag
     2761 gatacaggaa tccctgctga catctggcga ttctatctgc tatacattcg gcctgagggc
     2821 caggacagtg ccttctcctg gacagacttg ttgattaaaa acaattctga gctgctcaac
     2881 aacctgggca acttcatcaa cagagctggc atgtttgttt ctaagttttt tggcggttgt
     2941 gtgcctgaga tggcgctaac ccctgatgac agacgcctgg tggcccatgt ctcttgggaa
     3001 ctccagcact atcaccagct gttggaaaag gttcggatcc gggatgcctt gcgcagtatc
     3061 ctcaccatat ctcgccatgg caaccaatac attcaagtga atgagccctg gaaacggatt
     3121 aaaggtggtg agatggacag gcagcgggca ggcacagtga caggcatggc agtgaacatg
     3181 gctgccttgc tgtctgtcat gctgcagcca tacatgccca cagtcagctc taccatccag
     3241 acccagctgc agctcccacc tgcagcctgc cgcatccttg ccacaagctt catttgtacc
     3301 ttgccagcag gccaccgaat tggcacagtc agtcctttgt tccaaaaact ggaaaatgac
     3361 cagattgaaa atttgaggca gcgctttgga gggggtcagg ctaaaggctc ccccaagcca
     3421 gcagctgtgg aggcagttac agcagcaggc tcgcagcaca tacaaacgct gacggatgag
     3481 gtgaccaagc agggcaacgt cgtccgggaa ctgaaagcac agaaggcaga caagaaccag
     3541 gttgctgcag aggtggctaa actcttggat ctaaagaaac agttggcttt ggctgagggg
     3601 aaacccattg aaactcctaa aggcaagaag aaaaagtgat aactcgagtc tagagggccc
     3661 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3721 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3781 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3841 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3901 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3961 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4021 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4081 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4141 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4201 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4261 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4321 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4381 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4441 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4501 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4561 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4621 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4681 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4741 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4801 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4861 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4921 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4981 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5041 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5101 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5161 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5221 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5281 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5341 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5401 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5461 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5521 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5581 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5641 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5701 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5761 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5881 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5941 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6001 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6061 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6121 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6181 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6241 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6841 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
