LOCUS       Exported                6307 bp ds-DNA     circular SYN 22-MAR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6307)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 22, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6307
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNApolymerase,
                     with embedded lac operator"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    115..192
                     /label=14His tag
     exon            193..483
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
     CDS             484..2499
                     /codon_start=1
                     /label=nsp10-14 fusion protein
                     /translation="AGNATEVPANSTVLSFCAFAVDAAKAYKDYLASGGQPITNCVKML
                     CTHTGTGQAITVTPEANMDQESFGGASCCLYCRCHIDHPNPKGFCDLKGKYVQIPTTCA
                     NDPVGFTLKNTVCTVCGMWKGYGCSCDQLREPMLQGGSGGSAENVTGLFKDCSKVITGL
                     HPTQAPTHLSVDTKFKTEGLCVDIPGIPKDMTYRRLISMMGFKMNYQVNGYPNMFITRE
                     EAIRHVRAWIGFDVEGCHATREAVGTNLPLQLGFSTGVNLVAVPTGYVDTPNNTDFSRV
                     SAKPPPGDQFKHLIPLMYKGLPWNVVRIKIVQMLSDTLKNLSDRVVFVLWAHGFELTSM
                     KYFVKIGPERTCCLCDRRATCFSTASDTYACWHHSIGFDYVYNPFMIDVQQWGFTGNLQ
                     SNHDLYCQVHGNAHVASCDAIMTRCLAVHECFVKRVDWTIEYPIIGDELKINAACRKVQ
                     HMVVKAALLADKFPVLHDIGNPKAIKCVPQADVEWKFYDAQPCSDKAYKIEELFYSYAT
                     HSDKFTDGVCLFWNCNVDRYPANSIVCRFDTRVLSNLNLPGCDGGSLYVNKHAFHTPAF
                     DKSAFVNLKQLPFFYYSDSPCESHGKQVVSDIDYVPLKSATCITRCNLGGAVCRHHANE
                     YRLYLDAYNMMISAGFSLWVYKQFDTYNLWNTFTRLQ"
     misc_feature    484..900
                     /label=coNsp10
     misc_feature    901..918
                     /label=GGSGGS linker
     misc_feature    919..2499
                     /label=coNsp14
     terminator      2549..2643
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2704..3519
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3918..5000)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator 
                     toinhibit 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"
     promoter        complement(5001..5078)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in 
                     thepromoter boosts expression of the lacI gene about 
                     10-fold."
     rep_origin      complement(5596..6184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatgagc
      121 aagcatcacc atcattcagg ccatcaccat accggacacc accatcattc aggcagtcat
      181 caccataccg gatcggactc agaagtcaat caagaagcta agccagaggt caagccagaa
      241 gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga gatcttcttc
      301 aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa aagacagggt
      361 aaggaaatgg actccttaag attcttgtac gacggtatta gaattcaagc tgatcagacc
      421 cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga acagattggt
      481 ggcgcaggta atgcaaccga agttccggca aatagcaccg ttctgagctt ttgtgcattt
      541 gcagttgatg cagcaaaagc ctataaagac tatctggcaa gcggtggtca gccgattacc
      601 aattgtgtta aaatgctgtg tacccatacc ggcaccggtc aggcaattac cgttacaccg
      661 gaagcaaata tggatcaaga aagctttggt ggtgcaagct gttgtctgta ttgtcgttgt
      721 catattgatc acccgaatcc gaaaggtttt tgtgatctga aaggtaagta tgtgcagatt
      781 ccgaccacct gtgcaaatga tccggttggt tttaccctga aaaataccgt ttgcaccgtt
      841 tgtggtatgt ggaaaggtta tggttgtagc tgtgatcagc tgcgtgaacc gatgctgcag
      901 ggtggtagcg gtggtagtgc agaaaatgtt accggtctgt ttaaagattg cagcaaagtt
      961 attacaggtc tgcatccgac acaggcaccg acacatctga gcgttgatac caaattcaaa
     1021 accgaaggtc tgtgtgttga tattccgggt attccgaaag atatgaccta tcgtcgtctg
     1081 attagcatga tgggcttcaa aatgaattat caggtgaatg gctacccgaa catgtttatt
     1141 acccgtgaag aagcaattcg tcacgttcgt gcatggattg gttttgatgt tgaaggttgt
     1201 catgcaacac gtgaagcagt tggcaccaat ctgccgctgc agctgggttt tagcaccggt
     1261 gttaatctgg ttgcagttcc gaccggttat gttgataccc cgaataacac cgattttagc
     1321 cgtgttagcg caaaaccgcc tccgggtgat cagtttaaac atctgattcc gctgatgtat
     1381 aaaggtctgc cgtggaatgt tgtgcgtatc aaaattgttc agatgctgag cgataccctg
     1441 aaaaatctga gcgatcgtgt tgtttttgtt ctgtgggcac atggttttga actgaccagc
     1501 atgaaatact tcgtgaaaat tggtccggaa cgtacctgtt gtctgtgtga tcgtcgtgca
     1561 acctgtttta gtaccgcaag cgatacctat gcatgttggc atcatagcat tggcttcgat
     1621 tatgtgtata acccgtttat gattgatgtt cagcagtggg gttttaccgg caatctgcag
     1681 agcaatcatg atctgtattg tcaggttcat ggtaatgcac atgttgcaag ctgtgatgca
     1741 attatgaccc gttgtctggc agttcatgaa tgctttgtta aacgtgttga ttggaccatc
     1801 gaatatccga ttattggtga tgagctgaaa attaacgcag cctgtcgtaa agttcagcac
     1861 atggtggtta aagcagcact gctggcagat aaatttccgg ttctgcatga tattggtaac
     1921 ccgaaagcaa ttaaatgtgt tccgcaggca gatgtggaat ggaaattcta tgatgcacag
     1981 ccgtgtagcg ataaagccta taaaatcgaa gaactgttct atagctatgc gacccatagc
     2041 gacaaattta ccgatggtgt ttgtctgttt tggaattgca atgttgatcg ctatccggca
     2101 aatagcattg tttgccgttt tgatacccgt gttctgagca atctgaatct gcctggttgt
     2161 gatggtggta gcctgtatgt taacaaacat gcatttcata cccctgcctt tgataaaagc
     2221 gcatttgtga atctgaaaca gctgccgttt ttctattata gcgatagccc gtgtgaaagc
     2281 catggtaaac aggtggttag cgatattgat tatgttccgc tgaaaagcgc aacctgcatt
     2341 acccgttgta atcttggtgg tgcagtttgt cgtcatcatg caaatgaata tcgtctgtat
     2401 ctggatgcct acaacatgat gattagcgca ggttttagcc tgtgggtgta taaacagttt
     2461 gatacctata atctgtggaa cacctttaca cgtctgcagt aataagcggc cgcacatcac
     2521 catcactaag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
     2581 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     2641 aatccaagct agccatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     2701 gttatgagcc atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg
     2761 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2821 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2881 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2941 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcttggtt actcacgact
     3001 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     3061 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     3121 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     3181 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     3241 aaagaaatgc acaaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     3301 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     3361 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     3421 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     3481 aaattgcagt ttcatttgat gctcgatgag tttttctaag aattaattca tgggcaaata
     3541 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     3601 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     3661 gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg gggtaatgac
     3721 tctctagctt gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     3781 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ctctagatta
     3841 cgtgcagtcg atgataagct gtcaaacatg agaattgtgc ctaatgagtg agctaactta
     3901 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3961 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggggc cagggtggtt
     4021 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     4081 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     4141 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     4201 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4261 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     4321 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4381 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4441 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4501 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4561 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4621 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4681 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4741 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     4801 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     4861 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     4921 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     4981 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     5041 gccataccgc gaaaggtttt gcaccattcg atggtgtcgg aatttcgggc agcgttgggt
     5101 cctggccacg ggtgcgcatg atctagagct gcctcgcgcg tttcggtgat gacggtgaaa
     5161 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5221 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     5281 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     5341 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5401 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5461 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5521 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5581 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5641 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5701 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5761 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5821 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5881 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5941 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6001 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6061 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6121 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6241 ttaagggatt ttggtcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6301 tcttcac
//
