LOCUS       sequence-331999-        5118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Nsp3 Y domain (residues 1844-1945).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5118)
  TITLE     Savchenko Lab Plasmids
REFERENCE   2  (bases 1 to 5118)
  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_3
FEATURES             Location/Qualifiers
     source          1..5118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             73..90
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             115..135
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             514..531
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(584..602)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      598..645
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(769..788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        955..1058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1059..1127,1128..1919)
                     /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(1277..1296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2090..2678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2579..2598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2832..2849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2929..2948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3104..4186)
                     /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     4148..4167
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4187..4264)
                     /gene="lacI"
                     /label=lacI promoter
     tRNA            4404..4479
                     /gene="ileX"
                     /product="E. coli tRNA-Ile recognizing AUA"
                     /label=ileX
     primer_bind     4628..4647
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     tRNA            4749..4825
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
     primer_bind     join(5100..5118,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5100..5118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tgcaccatca tcatcatcat tcttctggtg tagatctggg taccgagaac
      121 ctgtacttcc aatccaatgc cgacatcgaa gttaccggtg atagctgcaa caactatatg
      181 ctgacctaca acaaagtgga gaacatgacc ccgagagatt taggtgcctg catcgattgt
      241 tctgcacgcc atattaatgc acaagtggca aaaagccaca acattgccct gatctggaac
      301 gtgaaggact tcatgagcct gagtgaacaa ctgcgtaaac agattcgttc tgcagccaag
      361 aagaacaatc tgccgttcaa actgacctgt gcaacaacac gtcaggtggt taatgttgtt
      421 accaccaaga ttgccctgaa aggtggttaa cattggaagt ggataacgga tccgaattcg
      481 agctccgtcg acaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg
      541 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
      601 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
      661 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      721 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      781 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      841 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      901 gtgatggttc acattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
      961 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1021 ccctgataaa tgcttcaata atttgaaaaa ggaagagtat gagtattcaa catttccgtg
     1081 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1141 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1201 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1261 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1321 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1381 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1441 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1501 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1561 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     1621 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1681 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1741 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1801 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1861 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1921 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1981 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2041 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2221 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2701 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2761 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2821 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     2881 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     2941 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg tagtcatgcc
     3001 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     3061 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     3121 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3181 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     3241 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     3301 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     3361 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     3421 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     3481 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     3541 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     3601 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     3661 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     3721 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     3781 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     3841 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     3901 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     3961 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     4021 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     4081 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     4141 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     4201 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     4261 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     4321 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcggatt gcgacacgga
     4381 gttactttat aatccgctac catggcccct tagctcagtg gttagagcag gcgactcata
     4441 atcgcttggt cgctggttca agtccagcag gggccaccag atatagcaaa ggctgacgag
     4501 aaatcgtcag cctttttgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     4561 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     4621 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggcgagat
     4681 caaaagccat tgactcagca agggttgacc gtataattca cgcgattaca ccgcattgcg
     4741 gtatcaacgc gcccttagct cagttggata gagcaacgac cttctaagtc gtgggccgca
     4801 ggttcgaatc ctgcagggcg cgccattaca attcaatcag ttacgccttc tttatatcct
     4861 ccataattcc agagtgggac atatttggga cattatcacc aaaaatgtcg tctattttcc
     4921 tcgcatgctc tgtcaaatga ttaggcgcaa ggtgagcata cctacgaacc atttctatgg
     4981 actcccatcc gcccatttcc tgaagcactg ataatgggac gcctgactga atccaccggt
     5041 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt
     5101 aatacgactc actatagg
//
