LOCUS       Exported                5849 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression of Nsp15.
ACCESSION   .
VERSION     .
KEYWORDS    pMCSG53_Nsp15
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5849)
  TITLE     Savchenko Lab Plasmids
REFERENCE   2  (bases 1 to 5849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5849
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1264..1281
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1334..1352)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1348..1395
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1519..1538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1705..1808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1809..2669
                     /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(2027..2046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2840..3428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3329..3348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3582..3599
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3679..3698)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3854..4936)
                     /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     4898..4917
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4937..5014)
                     /gene="lacI"
                     /label=lacI promoter
     tRNA            5154..5229
                     /gene="ileX"
                     /product="E. coli tRNA-Ile recognizing AUA"
                     /label=ileX
     primer_bind     5378..5397
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     tRNA            5499..5575
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
      121 agatctgggt accgagaacc tgtacttcca atccaatatg tcactagaaa atgtcgcttt
      181 taacgtagtt aacaaaggcc actttgacgg ccagcagggt gaagtgccgg ttagcatcat
      241 taacaatacc gtgtacacca aagtggacgg cgtcgatgtt gagctgttcg agaacaagac
      301 caccttgcca gtaaatgtgg cattcgagct gtgggcgaaa cgcaatatta agccggttcc
      361 ggaagtgaaa attctgaaca acctgggtgt ggacatcgcg gctaatacgg tgatctggga
      421 ttataagcgt gatgctccgg cacatattag caccattggc gtgtgtagca tgaccgacat
      481 tgccaaaaag ccgaccgaaa ccatttgcgc gccactgacc gtttttttcg acggccgtgt
      541 tgatggccag gttgacctgt ttagaaatgc tcgtaacggt gtgctgatta ccgagggtag
      601 cgttaaaggt cttcaaccgt ccgttggtcc gaagcaggca agcctcaacg gtgttaccct
      661 tatcggcgag gcggtgaaga cccagtttaa ctactacaaa aaagtcgacg gcgttgtgca
      721 gcaactgccg gaaacgtact ttacccagag ccgtaacctg caagaattta aaccgcgctc
      781 acagatggaa atcgacttcc tggagttagc catggatgaa ttcatcgaac gttataagct
      841 ggaaggctac gcgttcgagc acatcgtgta tggtgacttc agtcattctc aactgggtgg
      901 tctccacctg ctgatcggtt tggcgaagcg ctttaaagag tccccgtttg agttggagga
      961 cttcatcccg atggattcta cagttaagaa ctacttcatt actgacgcgc aaaccggatc
     1021 ctctaagtgc gtgtgcagcg tcatcgattt attgctggat gatttcgttg aaatcatcaa
     1081 aagccaagat ctgagcgtcg ttagcaaagt cgtgaaagta acgattgact acactgaaat
     1141 ctcgttcatg ttgtggtgta aagatggtca tgtcgagacg ttttatccga aactgcaata
     1201 aattggaagt ggataacgga tccgaattcg agctccgtcg acaagcttgc ggccgcactc
     1261 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag
     1321 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1381 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
     1441 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1501 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1561 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1621 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acattaacgc ttacaattta
     1681 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atttgaaaaa
     1801 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1861 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1921 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1981 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2041 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2101 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2161 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2221 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2281 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2341 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2401 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2461 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2521 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2581 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2641 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2701 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2761 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2821 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2881 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2941 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3001 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3061 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3121 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3181 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3241 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3301 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3361 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3421 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3481 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3541 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3601 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3661 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3721 ccgctatcgc tacgtgactg tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     3781 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     3841 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3901 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     3961 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4021 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4081 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     4141 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4201 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     4261 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     4321 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     4381 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     4441 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     4501 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     4561 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     4621 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     4681 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     4741 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     4801 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     4861 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     4921 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     4981 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5041 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5101 aggaatggtg catgcggatt gcgacacgga gttactttat aatccgctac catggcccct
     5161 tagctcagtg gttagagcag gcgactcata atcgcttggt cgctggttca agtccagcag
     5221 gggccaccag atatagcaaa ggctgacgag aaatcgtcag cctttttgca tgcaaggaga
     5281 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     5341 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5401 cagcaaccgc acctgtggcg ccggcgagat caaaagccat tgactcagca agggttgacc
     5461 gtataattca cgcgattaca ccgcattgcg gtatcaacgc gcccttagct cagttggata
     5521 gagcaacgac cttctaagtc gtgggccgca ggttcgaatc ctgcagggcg cgccattaca
     5581 attcaatcag ttacgccttc tttatatcct ccataattcc agagtgggac atatttggga
     5641 cattatcacc aaaaatgtcg tctattttcc tcgcatgctc tgtcaaatga ttaggcgcaa
     5701 ggtgagcata cctacgaacc atttctatgg actcccatcc gcccatttcc tgaagcactg
     5761 ataatgggac gcctgactga atccaccggt gatgccggcc acgatgcgtc cggcgtagag
     5821 gatcgagatc gatctcgatc ccgcgaaat
//
