LOCUS       sequence-252278-        8093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express INTS3 full-length in E. coli with a GST tag (N-terminal).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8093)
  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 8093)
  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: pGEX-6P-1-INTS3-FL
FEATURES             Location/Qualifiers
     source          1..8093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(540..545,546..563,564..570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    578..594
                     /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)."
     primer_bind     602..618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     602..618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             614..1141
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(634..651)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(634..650)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(643..665)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1243..1248,1249..1264,1265..1271)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1279..1295
                     /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             1318..1971
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1929..1951
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1978..2001
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5203..5225)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5370..5388)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5456..5560
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5561..5629,5630..6421)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5779..5798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6592..7180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7081..7100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7334..7351
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7424..7501
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(7502..8093,1..491)
                     /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     complement(7521..7540)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
       61 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
      121 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
      181 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
      241 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
      301 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
      361 tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct
      421 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
      481 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
      541 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      601 acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga
      661 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      721 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      781 tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga
      841 gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta
      901 cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc
      961 cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca
     1021 ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg
     1081 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     1141 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     1201 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg
     1261 ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg
     1321 tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg
     1381 gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg
     1441 cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt
     1501 gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg
     1561 ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat
     1621 attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat
     1681 tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca
     1741 tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt
     1801 gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa
     1861 cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg
     1921 cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg
     1981 gaagttctgt tccaggggcc cctgggatcc atggagttgc agaagggaaa aggggcggca
     2041 gcagcagcag ctgcttcggg agcagcggga ggtggaggag gaggagcggg agcaggagcc
     2101 ccaggagggg ggaggctgct actttcaacc agtttggatg ccaaggatga gttagaggag
     2161 agattggaaa ggtgtatgag cattgtgaca tcgatgactg ctggtgtctc ggagagagaa
     2221 gccaatgatg ccctcaatgc gtatgtgtgc aaaggcctcc cccagcatga agaaatctgc
     2281 ctgggcctgt ttactctcat cctcactgaa cctgcccaag cccagaagtg ttaccgggac
     2341 ttagctctgg tgagtcgtga tggcatgaat attgtcctga ataaaatcaa ccagatactt
     2401 atggagaagt acctgaagct gcaggatacc tgccgtactc agttggtgtg gttggtacgg
     2461 gaactggtga agagtggggt tctgggagcc gatggtgttt gtatgacgtt tatgaagcag
     2521 attgcaggtg gagatgttac agccaaaaat atctggttgg cagaaagtgt tctggatatc
     2581 ctgacagagc aaagggagtg ggtcctgaag agcagcatcc tcattgccat ggctgtttac
     2641 acgtacctcc gcctcatcgt ggaccaccat gggactgccc agctccaggc cctgcgacag
     2701 aaggaagtag acttctgcat ctcactgctt cgggaacggt tcatggaatg tctgatgatt
     2761 ggtcgggatc tcgtaagact acttcagaat gttgctagga taccagaatt tgaactgctt
     2821 tggaaagata ttatccataa tcctcaggcc ttgagtcctc agttcacagg tatcctacag
     2881 cttcttcagt caagaacatc ccgaaaattc ctagcatgtc gtctaacccc ggacatggag
     2941 actaaactcc tcttcatgac atcccgggtg cgatttggtc aacaaaagcg ataccaagat
     3001 tggttccagc gccagtacct gtcaactcca gatagtcagt ctctgcgctg tgacctcatt
     3061 cgctacatct gtggggtagt ccacccttct aatgaagtac tgagttcaga tatcttgccc
     3121 cggtgggcca tcattggttg gctcctgaca acgtgcacgt caaatgtcgc tgcctccaat
     3181 gccaagctgg ctttgtttta tgactggctg ttctttagtc cagacaagga tagcattatg
     3241 aacatagaac cagccatcct ggtcatgcac cactccatga agccccaccc agccatcact
     3301 gccacactcc tggacttcat gtgccgcatc attcccaact tctatccacc attggagggc
     3361 cacgtgcggc agggtgtctt ttcctccctc aaccacattg tggagaaacg ggtcttggca
     3421 cacctagctc ccctgtttga caaccctaag ttggataagg agctgcgggc aatgctgaga
     3481 gagaagtttc ctgagttctg cagctcaccc tccccacctg tggaagtcaa aattgaggag
     3541 ccagtttcca tggagatgga caaccatatg tcggataagg atgagagttg ctatgacaat
     3601 gcagaggcag ccttcagtga cgatgaagag gatctcaaca gcaaaggaaa gaagagggag
     3661 tttcgcttcc accctatcaa ggagacagtt gtggaggagc cagttgatat caccccttac
     3721 cttgaccagt tggatgagtc cctgagggac aaagtactcc agctacagaa ggggagtgat
     3781 acggaggccc agtgtgaggt catgcaggaa attgtggacc aggtcctgga ggaagacttt
     3841 gactcggagc agctgtctgt ccttgcttcc tgcctacagg agctcttcaa ggcccacttt
     3901 cgaggggagg tcctgcctga ggagattact gaggagtccc tggaggagtc tgtaggaaag
     3961 cctctgtacc taatatttag gaacctatgt cagatgcagg aagacaacag cagcttctct
     4021 ctacttctag accttctctc cgagctatat cagaagcagc ccaagattgg ctaccacctg
     4081 ctctactacc tgagggccag caaagccgcc gcagggaaga tgaacctgta cgagtcattt
     4141 gcccaggcta cccagctggg cgatctgcac acctgcctga tgatggacat gaaggcctgc
     4201 caggaggacg atgtgcggct cctgtgccac ctcacgccct ccatctacac agagtttcca
     4261 gatgaaacct tgaggagcgg agagctgctg aacatgatcg tggctgttat tgactctgca
     4321 cagctccagg agctggtctg ccacgtgatg atgggtaacc tggttatgtt tcgaaaagac
     4381 tcagttctca acatactcat tcagagccta gactgggaga cctttgagca gtattgtgcc
     4441 tggcagctct ttctggccca caatattccc ctggagacca taatccccat cctgcagcac
     4501 ctcaaataca aggagcaccc agaggccctg tcctgcctac tgcttcaact ccgaagagaa
     4561 aagcccagcg aggagatggt gaagatggtg ctgagccggc cctgccatcc tgacgaccag
     4621 ttcaccacca gcatcctgcg gcactggtgc atgaaacatg acgagctgct ggccgagcac
     4681 atcaagtccc tgctcatcaa gaacaacagc ctgcctcgca agagacagag cctgaggagc
     4741 tctagcagca agctggccca gctgactctg gagcagatcc tggagcactt ggacaatctg
     4801 cggctcaacc tgaccaacac caagcagaac ttttttagcc agacgccaat tctccaggcg
     4861 ctgcagcatg tccaagcgag ctgtgacgaa gcccacaaga tgaaattcag tgatctcttc
     4921 tccctggcgg aggaatatga ggactcttcc accaagccac ccaagagccg gcgaaaagca
     4981 gctctgtcca gccctcgaag tcgaaagaat gccacacagc cccccaatgc cgaagaagag
     5041 tcgggctcca gcagtgcttc agaagaggaa gacacgaaac cgaagcctac caagcggaaa
     5101 cgaaaagggt cctctgcagt gggctctgac agtgactctc gagcggccgc atcgtgactg
     5161 actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5221 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5281 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     5341 cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5401 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5461 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5521 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5581 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5641 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5701 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5761 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     5821 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5881 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5941 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6001 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6061 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     6121 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6181 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6241 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6301 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6361 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6421 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6481 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6541 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6601 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6661 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6721 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6781 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6841 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6901 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6961 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7021 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7081 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7141 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7201 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7261 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7321 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     7381 tctccttacg catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg
     7441 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     7501 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     7561 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     7621 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     7681 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     7741 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     7801 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     7861 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     7921 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     7981 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     8041 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtc
//
