LOCUS       sequence-304475-        7181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct for producing Avi tagged Arginyl-tRNA 
            synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7181)
  TITLE     Graslund lab plasmids
REFERENCE   2  (bases 1 to 7181)
  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: RARS_pNIC-Bio3
FEATURES             Location/Qualifiers
     source          1..7181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     969..988
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1008..1085)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1291..1310
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1394..1413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1394..1412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1413..1437
                     /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             1452..1474
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1485..1502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1527..1547
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3342..3386
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3439..3456
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3509..3527)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3523..3570
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3607..4062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3694..3713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3904..3925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4154..4969)
                     /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(4154..4969)
                     /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     4877..4896
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4961..4980
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5091..5679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5580..5599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5833..5850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5865..6004
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5930..5949)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6090..6112
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6106..6297)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7106..7181,1..1007))
                     /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
                     EACKAAVHNLLAQRVSGLIINYQLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
       61 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      121 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
      181 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
      241 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
      301 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
      361 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
      421 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
      481 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
      541 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
      601 acagcaatgg catcctggtc atccagctga tagttaatga tcagcccact gacgcgttgc
      661 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
      721 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
      781 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
      841 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
      901 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
      961 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1021 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     1081 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     1141 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     1201 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     1261 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     1321 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     1381 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     1441 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaccat catcatcatc
     1501 attcttctgg tgtagatctg ggtaccgaga acctgtactt ccaatccatg actaaaaata
     1561 tgattaacat tattagccgc ctacaagagg tctttggtca tgcaattaag gctgcatatc
     1621 cagatttgga aaatcctcct ctgctagtga caccaagtca gcaggccaag tttggggact
     1681 atcagtgtaa tagtgctatg ggtatttctc agatgctcaa aaccaaggaa cagaaagtta
     1741 atccaagaga aattgctgaa aacattacca aacacctccc agacaatgaa tgtattgaaa
     1801 aagttgaaat tgctggtcct ggttttatta atgtccactt aagaaaggat tttgtatcag
     1861 aacaattgac cagtcttcta gtgaatggag ttcaactacc tgctctggga gagaataaaa
     1921 aggttatagt tgacttttcc tcccctaata tagctaaaga gatgcatgta ggccacctga
     1981 ggtcaactat cataggagag agtataagcc gcctctttga atttgcaggg tatgacgtgc
     2041 tcaggttaaa tcatgtagga gactggggga cccagtttgg catgctcatc gctcacctgc
     2101 aagacaaatt tccagattat ctaacagttt cacctcctat tggggatctt caggtctttt
     2161 ataaggaatc taagaagagg tttgatactg aggaggaatt taagaagcga gcatatcagt
     2221 gtgtagttct gctccagggt aaaaacccag atattacaaa agcttggaag cttatctgtg
     2281 atgtctcccg ccaagagtta aataaaatct atgatgcatt ggacgtctct ttaatagaga
     2341 gaggggaatc cttctatcaa gataggatga atgatattgt aaaggaattt gaagatagag
     2401 gatttgtgca ggtggatgat ggcagaaaga ttgtatttgt cccagggtgt tccataccat
     2461 taaccatagt aaaatcagat ggaggttata cctatgatac atctgacctg gctgctatta
     2521 aacaaagact atttgaggaa aaagcagata tgattatcta tgttgtggac aatggacaat
     2581 ctgtgcactt ccagacaata tttgctgctg ctcaaatgat tggttggtat gaccctaaag
     2641 taactcgagt cttccatgct ggatttggtg tggtgctagg ggaagacaag aaaaagttta
     2701 aaacacgttc gggtgaaaca gtgcgcctca tggatcttct gggagaagga ctaaaacgat
     2761 ccatggacaa gttgaaggaa aaagaaagag acaaggtctt aactgcagag gaattgaatg
     2821 ctgctcagac atccgttgca tatggctgca tcaaatatgc tgacctttcc cataaccggt
     2881 tgaatgacta catcttctcc tttgacaaaa tgctagatga cagaggaaat acagctgctt
     2941 acttgttgta tgccttcact agaatcaggt ctattgcacg tctggccaat attgatgaag
     3001 aaatgctcca aaaagctgct cgagaaacca agattctttt ggatcatgag aaggaatgga
     3061 aactaggccg gtgcatttta cggttccctg agattctgca aaagatttta gatgacttat
     3121 ttctccacac tctctgtgat tatatatatg agctggcaac tgctttcaca gagttctatg
     3181 atagctgcta ctgtgtggag aaagatagac agactggaaa aatattgaag gtgaacatgt
     3241 ggcgtatgct gctatgtgaa gcagtagctg ctgtcatggc caaggggttt gatatcctgg
     3301 gaataaaacc tgtccaaagg atgagcagta aaggtggata cggcctgaat gatatctttg
     3361 aagcgcagaa gattgaatgg catgaatgat aaggatccga attcgagctc cgtcgacaag
     3421 cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     3481 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     3541 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     3601 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3661 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3721 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3781 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3841 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3901 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3961 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4021 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     4081 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4141 catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     4201 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     4261 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     4321 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     4381 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     4441 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     4501 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     4561 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     4621 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     4681 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     4741 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     4801 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     4861 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     4921 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg
     4981 tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag
     5041 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5101 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5161 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5221 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5281 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5341 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5401 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5461 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5521 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5581 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5641 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5701 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5761 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5821 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     5881 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     5941 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     6001 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     6061 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     6121 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     6181 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     6241 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     6301 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     6361 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     6421 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     6481 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     6541 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     6601 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     6661 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     6721 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
     6781 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     6841 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     6901 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     6961 gataaagaag acagtcataa gtgcggcgac gatagtcatg tcccgcgccc accggaagga
     7021 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     7081 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7141 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc g
//
