LOCUS       sequence-304479-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct for producing Avi tagged Tyrosyl-tRNA 
            synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  TITLE     Graslund lab plasmids
REFERENCE   2  (bases 1 to 6894)
  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: YARS_pNIC-CTB10H
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     971..990
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1010..1087)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1293..1312
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1396..1415
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1396..1414
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1415..1439
                     /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             3070..3090
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3097..3141
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3142..3171
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3224..3242)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3238..3285
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3322..3777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3409..3428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3619..3640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3869..4684)
                     /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(3869..4684)
                     /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     4592..4611
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4676..4695
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4806..5394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5295..5314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5548..5565
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5580..5719
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5645..5664)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5805..5827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5821..6012)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6821..6894,1..1009))
                     /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"
ORIGIN
        1 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
       61 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
      121 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
      181 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
      241 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
      301 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
      361 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
      421 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
      481 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
      541 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
      601 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
      661 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
      721 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
      781 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
      841 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
      901 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
      961 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     1021 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     1081 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     1141 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     1201 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     1261 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     1321 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     1381 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     1441 ctctagaaat aattttgttt aaccttaaga aggagatata ctatggggga cgctcccagc
     1501 cctgaagaga aactgcacct tatcacccgg aacctgcagg aggttctggg ggaagagaag
     1561 ctgaaggaga tactgaagga gcgggaactt aaaatttact ggggaacggc aaccacgggc
     1621 aaaccacatg tggcttactt tgtgcccatg tcaaagattg cagacttctt aaaggcaggg
     1681 tgtgaggtaa caattctgtt tgcggacctc cacgcatacc tggataacat gaaagcccca
     1741 tgggaacttc tagaactccg agtcagttac tatgagaatg tgatcaaagc aatgctggag
     1801 agcattggtg tgcccttgga gaagctcaag ttcatcaaag gcactgatta ccagctcagc
     1861 aaagagtaca cactagatgt gtacagactc tcctccgtgg tcacacagca cgattccaag
     1921 aaggctggag ctgaggtggt aaagcaggtg gagcaccctt tgctgagtgg cctcttatac
     1981 cccggactgc aggctttgga tgaagagtat ttaaaagtag atgcccaatt tggaggcatt
     2041 gatcagagaa agattttcac ctttgcagag aagtacctcc ctgcacttgg ctattcaaaa
     2101 cgggtccatc tgatgaatcc tatggttcca ggattaacag gcagcaaaat gagctcttca
     2161 gaagaggagt ccaagattga tctccttgat cggaaggagg atgtgaagaa aaaactgaag
     2221 aaggccttct gtgagccagg aaatgtggag aacaatgggg ttctgtcctt catcaagcat
     2281 gtcctttttc cccttaagtc cgagtttgtg atcctacgag atgagaaatg gggtggaaac
     2341 aaaacctaca cagcttacgt ggacctggaa aaggactttg ctgctgaggt tgtacatcct
     2401 ggagacctga agaattctgt tgaagtcgca ctgaacaagt tgctggatcc aatccgggaa
     2461 aagtttaata cccctgccct gaaaaaactg gccagcgctg cctacccaga tccctcaaag
     2521 cagaagccaa tggccaaagg ccctgccaag aattcagaac cagaggaggt catcccatcc
     2581 cggctggata tccgtgtggg gaaaatcatc actgtggaga agcacccaga tgcagacagc
     2641 ctgtatgtag agaagattga cgtgggggaa gctgaaccac ggactgtggt gagcggcctg
     2701 gtacagttcg tgcccaagga ggaactgcag gacaggctgg tagtggtgct gtgcaacctg
     2761 aaaccccaga agatgagagg agtcgagtcc caaggcatgc ttctgtgtgc ttctatagaa
     2821 gggataaacc gccaggttga acctctggac cctccggcag gctctgctcc tggtgagcac
     2881 gtgtttgtga agggctatga aaagggccaa ccagatgagg agctcaagcc caagaagaaa
     2941 gtcttcgaga agttgcaggc tgacttcaaa atttctgagg agtgcatcgc acagtggaag
     3001 caaaccaact tcatgaccaa gctgggctcc atttcctgta aatcgctgaa aggggggaac
     3061 attagcgcag agaacctcta cttccaatcg ggtggcggtc tgaacgacat cttcgaggct
     3121 cagaaaatcg aatggcacga acatcatcat catcaccacc accaccacca ctgagatccg
     3181 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3241 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     3301 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3361 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3541 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3601 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3661 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3721 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     3781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3841 aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact
     3901 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     3961 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     4021 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     4081 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     4141 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     4201 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     4261 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     4321 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     4381 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     4441 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     4501 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     4561 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     4621 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     4681 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa
     4741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4921 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5461 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5521 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5581 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
     5641 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     5701 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5761 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5821 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     5881 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     5941 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     6001 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     6061 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     6121 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     6181 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     6241 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     6301 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     6361 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     6421 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     6481 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     6541 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     6601 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     6661 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     6721 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     6781 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6841 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gagg
//
