LOCUS       sequence-304476-        6852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct for producing Avi tagged Seryl-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  TITLE     Graslund lab plasmids
REFERENCE   2  (bases 1 to 6852)
  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: SARS_pNIC-CTB10H
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     350..369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     603..620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    635..774
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(700..719)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     860..882
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(876..1067)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1876..2958)
                     /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     2920..2939
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2959..3036)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3242..3261
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3345..3364
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3345..3363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3364..3388
                     /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)."
     CDS             4977..4997
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5004..5048
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5049..5078
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(5131..5149)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5145..5192
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5229..5684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5316..5335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5526..5547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5776..6591)
                     /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(5776..6591)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6499..6518
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6583..6602
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(6713..6852,1..449)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
       61 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
      121 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      181 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      241 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      301 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      361 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      421 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      481 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      541 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      601 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
      661 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
      721 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
      781 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      841 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      901 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
      961 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     1021 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     1081 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     1141 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     1201 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     1261 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     1321 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     1381 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     1441 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     1501 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     1561 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     1621 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     1681 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     1741 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     1801 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     1861 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     1981 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     2041 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     2101 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     2161 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     2221 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     2281 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     2341 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     2401 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     2461 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     2521 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     2581 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     2641 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     2701 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     2761 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     2821 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     2881 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     2941 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     3001 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     3061 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     3121 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     3181 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     3241 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     3301 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
     3361 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta accttaagaa
     3421 ggagatatac tatggtgctg gatctggatt tgtttcgggt ggataaagga ggggacccag
     3481 ccctcatccg agagacgcag gagaagcgct tcaaggaccc gggactagtg gaccagctgg
     3541 tgaaggcaga cagcgagtgg cgacgatgta gatttcgggc agacaacttg aacaagctga
     3601 agaacctatg cagcaagaca atcggagaga aaatgaagaa aaaagagcca gtgggagatg
     3661 atgagtctgt cccagagaat gtgctgagtt tcgatgacct tactgcagac gctttagcta
     3721 acctgaaagt ctcacaaatc aaaaaagtcc gactcctcat tgatgaagcc atcctgaagt
     3781 gtgacgcgga gcggataaag ttggaagcag agcggtttga gaacctccga gagattggga
     3841 accttctgca cccttctgta cccatcagta acgatgagga tgtggacaac aaagtagaga
     3901 ggatttgggg tgattgtaca gtcaggaaga agtactctca tgtggacctg gtggtgatgg
     3961 tagatggctt tgaaggcgaa aagggggccg tggtggctgg gagtcgaggg tacttcttga
     4021 agggggtcct ggtgttcctg gaacaggctc tcatccagta tgcccttcgc accttgggaa
     4081 gtcggggcta cattcccatt tataccccct ttttcatgag gaaggaggtc atgcaggagg
     4141 tggcacagct cagccagttt gatgaagaac tttataaggt gattggcaaa ggcagtgaaa
     4201 agtctgatga caactcctat gatgagaagt acctgattgc cacctcagag cagcccattg
     4261 ctgccctgca ccgggatgag tggctccggc cggaggacct gcccatcaag tatgctggcc
     4321 tgtctacctg cttccgtcag gaggtgggct cccatggccg tgacacccgt ggcatcttcc
     4381 gagtccatca gtttgagaag attgaacagt ttgtgtactc atcaccccat gacaacaagt
     4441 catgggagat gtttgaagag atgattacca ccgcagagga gttctaccag tccctgggga
     4501 ttccttacca cattgtgaat attgtctcag gttctttgaa tcatgctgcc agtaagaagc
     4561 ttgacctgga ggcctggttt ccgggctcag gagccttccg tgagttggtc tcctgttcta
     4621 attgcacgga ttaccaggct cgccggcttc gaatccgata tgggcaaacc aagaagatga
     4681 tggacaaggt ggagtttgtc catatgctca atgctaccat gtgcgccact acccgtacca
     4741 tctgcgccat cctggagaac taccagacag agaagggcat cactgtgcct gagaaattga
     4801 aggagttcat gccgccagga ctgcaagaac tgatcccctt tgtgaagcct gcgcccattg
     4861 agcaggagcc atcaaagaag cagaagaagc aacatgaggg cagcaaaaag aaagcagcag
     4921 caagagacgt caccctagaa aacaggctgc agaacatgga ggtcaccgat gctgcagaga
     4981 acctctactt ccaatcgggt ggcggtctga acgacatctt cgaggctcag aaaatcgaat
     5041 ggcacgaaca tcatcatcat caccaccacc accaccactg agatccggct gctaacaaag
     5101 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5161 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     5221 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5281 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5341 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5401 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5461 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5521 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5581 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5641 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     5701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5761 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     5821 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     5881 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     5941 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     6001 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     6061 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     6121 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     6181 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     6241 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     6301 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     6361 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     6421 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     6481 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6541 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     6601 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     6661 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6781 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6841 cgcagatacc aa
//
