LOCUS       sequence-252194-        6885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express INTS3 C-termini (514-1042 aa) in E. coli with N-terminal and
            C-terminal His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6885)
  AUTHORS   Vidhyasagar V, He Y, Guo M, Talwar T, Singh RS, Yadav M, Katselis G,
            Vizeacoumar FJ, Lukong KE, Wu Y
  TITLE     Biochemical characterization of INTS3 and C9ORF80, two subunits of 
            hNABP1/2 heterotrimeric complex in nucleic acid binding.
  JOURNAL   Biochem J. 2018 Jan 2;475(1):45-60. doi: 10.1042/BCJ20170351.
  PUBMED    29150435
REFERENCE   2  (bases 1 to 6885)
  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"; doi: "10"; journalName: 
            "Biochem J"; date: "2018-01-2- 2"; volume: "475"; issue: "1"; pages:
            "45-60"
COMMENT     Sequence Label: pET28a-INTS3-C
FEATURES             Location/Qualifiers
     source          1..6885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     144..163
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        144..162
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    163..187
                     /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             202..224
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             243..260
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             270..287
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1893..1910
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1963..1981)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1977..2024
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2061..2516
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2148..2167)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2358..2379
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2608..3423)
                     /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(2608..3423)
                     /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     3331..3350
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3415..3434
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3545..4133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4034..4053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4287..4304
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4319..4458
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4384..4403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4544..4566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4560..4751)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5560..6642)
                     /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     6604..6623
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6643..6720)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
       61 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
      121 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
      181 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
      241 gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat atggctagca
      301 tggagatgga caaccatatg tcggataagg atgagagttg ctatgacaat gcagaggcag
      361 ccttcagtga cgatgaagag gatctcaaca gcaaaggaaa gaagagggag tttcgcttcc
      421 accctatcaa ggagacagtt gtggaggagc cagttgatat caccccttac cttgaccagt
      481 tggatgagtc cctgagggac aaagtactcc agctacagaa ggggagtgat acggaggccc
      541 agtgtgaggt catgcaggaa attgtggacc aggtcctgga ggaagacttt gactcggagc
      601 agctgtctgt ccttgcttcc tgcctacagg agctcttcaa ggcccacttt cgaggggagg
      661 tcctgcctga ggagattact gaggagtccc tggaggagtc tgtaggaaag cctctgtacc
      721 taatatttag gaacctatgt cagatgcagg aagacaacag cagcttctct ctacttctag
      781 accttctctc cgagctatat cagaagcagc ccaagattgg ctaccacctg ctctactacc
      841 tgagggccag caaagccgcc gcagggaaga tgaacctgta cgagtcattt gcccaggcta
      901 cccagctggg cgatctgcac acctgcctga tgatggacat gaaggcctgc caggaggacg
      961 atgtgcggct cctgtgccac ctcacgccct ccatctacac agagtttcca gatgaaacct
     1021 tgaggagcgg agagctgctg aacatgatcg tggctgttat tgactctgca cagctccagg
     1081 agctggtctg ccacgtgatg atgggtaacc tggttatgtt tcgaaaagac tcagttctca
     1141 acatactcat tcagagccta gactgggaga cctttgagca gtattgtgcc tggcagctct
     1201 ttctggccca caatattccc ctggagacca taatccccat cctgcagcac ctcaaataca
     1261 aggagcaccc agaggccctg tcctgcctac tgcttcaact ccgaagagaa aagcccagcg
     1321 aggagatggt gaagatggtg ctgagccggc cctgccatcc tgacgaccag ttcaccacca
     1381 gcatcctgcg gcactggtgc atgaaacatg acgagctgct ggccgagcac atcaagtccc
     1441 tgctcatcaa gaacaacagc ctgcctcgca agagacagag cctgaggagc tctagcagca
     1501 agctggccca gctgactctg gagcagatcc tggagcactt ggacaatctg cggctcaacc
     1561 tgaccaacac caagcagaac ttttttagcc agacgccaat tctccaggcg ctgcagcatg
     1621 tccaagcgag ctgtgacgaa gcccacaaga tgaaattcag tgatctcttc tccctggcgg
     1681 aggaatatga ggactcttcc accaagccac ccaagagccg gcgaaaagca gctctgtcca
     1741 gccctcgaag tcgaaagaat gccacacagc cccccaatgc cgaagaagag tcgggctcca
     1801 gcagtgcttc agaagaggaa gacacgaaac cgaagcctac caagcggaaa cgaaaagggt
     1861 cctctgcagt gggctctgac agtgacctcg agcaccacca ccaccaccac tgagatccgg
     1921 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     1981 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     2041 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2101 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2161 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2221 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2281 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2341 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2401 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2461 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     2521 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2581 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
     2641 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     2701 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     2761 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     2821 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     2881 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     2941 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     3001 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     3061 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     3121 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     3181 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     3241 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     3301 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     3361 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     3421 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa
     3481 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3541 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3601 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3661 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4021 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4081 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4141 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4201 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4261 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4321 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     4381 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     4441 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4501 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4561 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     4621 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     4681 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4741 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     4801 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     4861 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     4921 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     4981 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     5041 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     5101 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     5161 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg
     5221 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     5281 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     5341 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     5401 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     5461 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     5521 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5581 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5641 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     5701 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     5761 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     5821 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     5881 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5941 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     6001 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     6061 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     6121 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     6181 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     6241 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     6301 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     6361 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     6421 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     6481 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     6541 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     6601 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     6661 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     6721 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     6781 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     6841 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaag
//
