LOCUS       sequence-331997-        5793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Nsp3 Y domain (residues 1619-1945).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5793)
  TITLE     Savchenko Lab Plasmids
REFERENCE   2  (bases 1 to 5793)
  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_2
FEATURES             Location/Qualifiers
     source          1..5793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      115..703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     604..623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     857..874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(954..973)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(1129..2211)
                     /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     2173..2192
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2212..2289)
                     /gene="lacI"
                     /label=lacI promoter
     tRNA            2429..2504
                     /gene="ileX"
                     /product="E. coli tRNA-Ile recognizing AUA"
                     /label=ileX
     primer_bind     2653..2672
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     tRNA            2774..2850
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
     primer_bind     3125..3144
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3125..3143
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3144..3168
                     /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             3183..3205
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3216..3233
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3258..3278
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4332..4349
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4402..4420)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4416..4463
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4587..4606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4773..4876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4877..4945,4946..5737)
                     /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(5095..5114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
       61 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
      121 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
      181 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
      241 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
      301 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
      361 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
      421 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
      481 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
      541 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
      601 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
      661 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
      721 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
      781 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
      841 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
      901 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
      961 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgtagtc
     1021 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     1081 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     1141 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1201 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     1261 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     1321 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     1381 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     1441 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     1501 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     1561 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     1621 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     1681 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     1741 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     1801 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     1861 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     1921 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     1981 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     2041 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     2101 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     2161 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     2221 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     2281 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     2341 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc ggattgcgac
     2401 acggagttac tttataatcc gctaccatgg ccccttagct cagtggttag agcaggcgac
     2461 tcataatcgc ttggtcgctg gttcaagtcc agcaggggcc accagatata gcaaaggctg
     2521 acgagaaatc gtcagccttt ttgcatgcaa ggagatggcg cccaacagtc ccccggccac
     2581 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     2641 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggc
     2701 gagatcaaaa gccattgact cagcaagggt tgaccgtata attcacgcga ttacaccgca
     2761 ttgcggtatc aacgcgccct tagctcagtt ggatagagca acgaccttct aagtcgtggg
     2821 ccgcaggttc gaatcctgca gggcgcgcca ttacaattca atcagttacg ccttctttat
     2881 atcctccata attccagagt gggacatatt tgggacatta tcaccaaaaa tgtcgtctat
     2941 tttcctcgca tgctctgtca aatgattagg cgcaaggtga gcatacctac gaaccatttc
     3001 tatggactcc catccgccca tttcctgaag cactgataat gggacgcctg actgaatcca
     3061 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct cgatcccgcg
     3121 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     3181 attttgttta actttaagaa ggagatatac atatgcacca tcatcatcat cattcttctg
     3241 gtgtagatct gggtaccgag aacctgtact tccaatccaa tgcctatgca aatggcggta
     3301 agggcttctg taaactgcac aactggaatt gcgtgaactg cgacaccttt tgtgcaggtt
     3361 caacctttat cagcgacgaa gttgcaagag atttaagcct gcagttcaaa cgcccgatta
     3421 atccgaccga tcagagtagc tacatcgttg atagcgttac cgtgaaaaat ggcagcatcc
     3481 acctgtattt cgacaaagcc ggccagaaaa cctatgaacg tcattctctg agccatttcg
     3541 tgaacctgga caatttacgc gccaacaaca ccaaaggtag tctgccgatt aacgtgatcg
     3601 tgtttgacgg taagagcaaa tgtgaagaga gcagcgcgaa atcagcaagc gtgtactata
     3661 gtcagctgat gtgtcagcct attctgttac tggatcaagc cctggttagc gatgtgggtg
     3721 atagtgcaga agttgccgtt aagatgtttg atgcctatgt gaacaccttc agcagcacct
     3781 tcaatgtgcc gatggagaaa ctgaagacct tagttgcaac cgcagaagca gaattagcca
     3841 agaacgtttc actggacaac gtgctgagca cctttatttc agcagcacgt cagggttttg
     3901 ttgacagcga tgttgaaacc aaagatgtgg tggagtgcct gaaattaagc catcagagcg
     3961 acatcgaagt taccggtgat agctgcaaca actatatgct gacctacaac aaagtggaga
     4021 acatgacccc gagagattta ggtgcctgca tcgattgttc tgcacgccat attaatgcac
     4081 aagtggcaaa aagccacaac attgccctga tctggaacgt gaaggacttc atgagcctga
     4141 gtgaacaact gcgtaaacag attcgttctg cagccaagaa gaacaatctg ccgttcaaac
     4201 tgacctgtgc aacaacacgt caggtggtta atgttgttac caccaagatt gccctgaaag
     4261 gtggttaaca ttggaagtgg ataacggatc cgaattcgag ctccgtcgac aagcttgcgg
     4321 ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
     4381 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     4441 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     4501 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4561 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4621 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4681 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac attaacgctt
     4741 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4801 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4981 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5041 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5101 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5161 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5221 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5281 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5341 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5401 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     5461 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5521 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5581 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5641 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5701 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5761 tatactttag attgatttaa aacttcattt tta
//
