LOCUS       sequence-304480-        6654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct for producing Avi tagged Tyrosyl-tRNA 
            synthetase 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6654)
  TITLE     Graslund lab plasmids
REFERENCE   2  (bases 1 to 6654)
  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: YARS2_pNIC-CTB10H
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     355..374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     608..625
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    640..779
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(705..724)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     865..887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(881..1072)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1881..2963)
                     /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     2925..2944
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2964..3041)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3247..3266
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3350..3369
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3350..3368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3369..3393
                     /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             4784..4804
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4811..4855
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4856..4885
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(4938..4956)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4952..4999
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5036..5491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5123..5142)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5333..5354
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5583..6398)
                     /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(5583..6398)
                     /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     6306..6325
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6390..6409
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(6520..6654,1..454)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
       61 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      121 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      181 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      241 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      301 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      361 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      421 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      481 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      541 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      601 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
      661 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
      721 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
      781 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      841 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      901 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
      961 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     1021 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     1081 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     1141 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     1201 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     1261 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     1321 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     1381 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     1441 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     1501 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     1561 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     1621 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     1681 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     1741 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     1801 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     1861 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     1981 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     2041 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     2101 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     2161 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     2221 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     2281 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     2341 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     2401 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     2461 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     2521 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     2581 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     2641 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     2701 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     2761 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     2821 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     2881 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     2941 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     3001 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     3061 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     3121 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     3181 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     3241 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     3301 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     3361 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacctt
     3421 aagaaggaga tatactatgg cccactcggg cgctcagggg ttactggcag cgcagaaggc
     3481 tcgaggtctg ttcaaggact tcttcccgga gacggggacg aaaatagagc tcccagagct
     3541 cttcgaccgt ggcacggcga gttttcccca aaccatttac tgtggcttcg accccacggc
     3601 agactcgctt catgtgggtc atctacttgc gctgctgggc ctgtttcatt tgcagcgagc
     3661 gggccacaac gtgatcgcgc tggtgggagg cgccacggcg cgcctgggag acccgagcgg
     3721 ccgtaccaag gaacgcgagg cgctggagac agagcgcgtg cgagccaacg cgcgagctct
     3781 gcgcctaggg cttgaggccc tggcggctaa tcaccagcag cttttcactg atgggcgctc
     3841 ctggggcagc ttcactgtgc tggacaactc ggcctggtac cagaagcagc acctggtgga
     3901 cttcctggcg gcagtggggg gtcacttccg catggggacg ctgctgagcc ggcagagcgt
     3961 gcagctgcgg ctcaagagcc ccgagggcat gagcttggcc gagttctttt accaggtgct
     4021 ccaggcctat gacttctatt acctcttcca gcgttatgga tgcagggtcc agctgggcgg
     4081 atctgatcaa ctaggcaaca tcatgtccgg atatgagttc atcaacaagt tgactggaga
     4141 agatgtattt ggaatcaccg ttcctctaat tacaagtaca actggagcaa agctgggaaa
     4201 gtctgctggc aacgctgttt ggctaaacag agataagaca tctccatttg aattgtatca
     4261 attctttgtc aggcaaccgg acgattcagt ggaaaggtac ctgaagctgt tcactttcct
     4321 gccccttcca gagattgatc atatcatgca gctgcatgtc aaagagccag aaaggcgggg
     4381 tcctcagaaa cgactggcag cagaagtaac aaagcttgtt catggacgag aaggattgga
     4441 ttctgctaaa aggtgtacac aagcccttta tcacagtagc atagatgcac tggaggtcat
     4501 gtctgatcag gagttaaaag agttgtttaa agaagctcca ttttctgaat tttttctcga
     4561 tcctggaaca agtgtcctag atacttgccg caaagcaaat gccattccag atggtccccg
     4621 agggtatcga atgataacag aaggcggagt cagcataaat caccaacaag taacaaatcc
     4681 tgagagtgtt ttaattgttg gacaacatat tctcaagaat ggactttcct tacttaaaat
     4741 aggaaaaaga aatttctaca ttataaaatg gcttcagttg gcagagaacc tctacttcca
     4801 atcgggtggc ggtctgaacg acatcttcga ggctcagaaa atcgaatggc acgaacatca
     4861 tcatcatcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     4921 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     4981 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     5041 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5101 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5161 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5221 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5281 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5341 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5401 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5461 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     5521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     5581 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     5641 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     5701 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     5761 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     5821 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     5881 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     5941 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     6001 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     6061 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     6121 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     6181 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     6241 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     6301 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     6361 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     6421 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     6481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agat
//
