LOCUS       sequence-331341-        5898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Nsp3 Y domain (residues 1584-1945).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  TITLE     Savchenko Lab Plasmids
REFERENCE   2  (bases 1 to 5898)
  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"
COMMENT     Sequence Label: pMCSG53_Nsp3_Y_1
FEATURES             Location/Qualifiers
     source          1..5898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     932..951
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(971..1048)
                     /gene="lacI"
                     /label=lacI promoter
     tRNA            1188..1263
                     /gene="ileX"
                     /product="E. coli tRNA-Ile recognizing AUA"
                     /label=ileX
     primer_bind     1412..1431
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     tRNA            1533..1609
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
     primer_bind     1884..1903
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1884..1902
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1903..1927
                     /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)."
     RBS             1942..1964
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1975..1992
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2017..2037
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3196..3213
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3266..3284)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3280..3327
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3451..3470)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3637..3740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3741..3809,3810..4601)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3959..3978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4772..5360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5261..5280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5514..5531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5611..5630)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(5786..5898,1..970))
                     /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"
ORIGIN
        1 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
       61 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
      121 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
      181 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
      241 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
      301 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
      361 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
      421 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
      481 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
      541 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
      601 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
      661 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
      721 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
      781 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
      841 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
      901 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
      961 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     1021 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     1081 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     1141 ggtgcatgcg gattgcgaca cggagttact ttataatccg ctaccatggc cccttagctc
     1201 agtggttaga gcaggcgact cataatcgct tggtcgctgg ttcaagtcca gcaggggcca
     1261 ccagatatag caaaggctga cgagaaatcg tcagcctttt tgcatgcaag gagatggcgc
     1321 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     1381 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     1441 ccgcacctgt ggcgccggcg agatcaaaag ccattgactc agcaagggtt gaccgtataa
     1501 ttcacgcgat tacaccgcat tgcggtatca acgcgccctt agctcagttg gatagagcaa
     1561 cgaccttcta agtcgtgggc cgcaggttcg aatcctgcag ggcgcgccat tacaattcaa
     1621 tcagttacgc cttctttata tcctccataa ttccagagtg ggacatattt gggacattat
     1681 caccaaaaat gtcgtctatt ttcctcgcat gctctgtcaa atgattaggc gcaaggtgag
     1741 catacctacg aaccatttct atggactccc atccgcccat ttcctgaagc actgataatg
     1801 ggacgcctga ctgaatccac cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     1861 gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     1921 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaccat
     1981 catcatcatc attcttctgg tgtagatctg ggtaccgaga acctgtactt ccaatccaat
     2041 gccgatggct gcaacagtag tacctgtatg atgtgctaca aacgtaatcg tgcaacacgc
     2101 gttgaatgta ccaccatcgt gaatggtgtg cgtcgtagct tctatgttta tgcaaatggc
     2161 ggtaagggct tctgtaaact gcacaactgg aattgcgtga actgcgacac cttttgtgca
     2221 ggttcaacct ttatcagcga cgaagttgca agagatttaa gcctgcagtt caaacgcccg
     2281 attaatccga ccgatcagag tagctacatc gttgatagcg ttaccgtgaa aaatggcagc
     2341 atccacctgt atttcgacaa agccggccag aaaacctatg aacgtcattc tctgagccat
     2401 ttcgtgaacc tggacaattt acgcgccaac aacaccaaag gtagtctgcc gattaacgtg
     2461 atcgtgtttg acggtaagag caaatgtgaa gagagcagcg cgaaatcagc aagcgtgtac
     2521 tatagtcagc tgatgtgtca gcctattctg ttactggatc aagccctggt tagcgatgtg
     2581 ggtgatagtg cagaagttgc cgttaagatg tttgatgcct atgtgaacac cttcagcagc
     2641 accttcaatg tgccgatgga gaaactgaag accttagttg caaccgcaga agcagaatta
     2701 gccaagaacg tttcactgga caacgtgctg agcaccttta tttcagcagc acgtcagggt
     2761 tttgttgaca gcgatgttga aaccaaagat gtggtggagt gcctgaaatt aagccatcag
     2821 agcgacatcg aagttaccgg tgatagctgc aacaactata tgctgaccta caacaaagtg
     2881 gagaacatga ccccgagaga tttaggtgcc tgcatcgatt gttctgcacg ccatattaat
     2941 gcacaagtgg caaaaagcca caacattgcc ctgatctgga acgtgaagga cttcatgagc
     3001 ctgagtgaac aactgcgtaa acagattcgt tctgcagcca agaagaacaa tctgccgttc
     3061 aaactgacct gtgcaacaac acgtcaggtg gttaatgttg ttaccaccaa gattgccctg
     3121 aaaggtggtt aacattggaa gtggataacg gatccgaatt cgagctccgt cgacaagctt
     3181 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
     3241 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     3301 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     3361 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3421 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3481 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3541 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacattaac
     3601 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3661 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3721 taatttgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3781 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3841 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3901 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3961 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4021 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4081 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4141 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4201 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4261 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     4321 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4381 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4441 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4501 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4561 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4621 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4681 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4741 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4801 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4861 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4921 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4981 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5041 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5101 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5161 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5221 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5281 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5341 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5401 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5461 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5521 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5581 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5641 ccagtataca ctccgctatc gctacgtgac tgtagtcatg ccccgcgccc accggaagga
     5701 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     5761 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5821 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     5881 gggtggtttt tcttttca
//
