LOCUS       sequence-246092-        8096 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 8096)
  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 8096)
  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..8096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(4146..4168)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4313..4331)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4399..4503
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4504..4572,4573..5364)
                     /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(4722..4741)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5535..6123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6024..6043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6277..6294
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6367..6444
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6445..7527
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(6464..6483)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    7540..7561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7576..7581,7582..7599,7600..7606)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7614..7630
                     /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     7638..7654
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7638..7654
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7650..8096,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7670..7687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7670..7686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7679..7701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atggagttgc
      961 agaagggaaa aggggcggca gcagcagcag ctgcttcggg agcagcggga ggtggaggag
     1021 gaggagcggg agcaggagcc ccaggagggg ggaggctgct actttcaacc agtttggatg
     1081 ccaaggatga gttagaggag agattggaaa ggtgtatgag cattgtgaca tcgatgactg
     1141 ctggtgtctc ggagagagaa gccaatgatg ccctcaatgc gtatgtgtgc aaaggcctcc
     1201 cccagcatga agaaatctgc ctgggcctgt ttactctcat cctcactgaa cctgcccaag
     1261 cccagaagtg ttaccgggac ttagctctgg tgagtcgtga tggcatgaat attgtcctga
     1321 ataaaatcaa ccagatactt atggagaagt acctgaagct gcaggatacc tgccgtactc
     1381 agttggtgtg gttggtacgg gaactggtga agagtggggt tctgggagcc gatggtgttt
     1441 gtatgacgtt tatgaagcag attgcaggtg gagatgttac agccaaaaat atctggttgg
     1501 cagaaagtgt tctggatatc ctgacagagc aaagggagtg ggtcctgaag agcagcatcc
     1561 tcattgccat ggctgtttac acgtacctcc gcctcatcgt ggaccaccat gggactgccc
     1621 agctccaggc cctgcgacag aaggaagtag acttctgcat ctcactgctt cgggaacggt
     1681 tcatggaatg tctgatgatt ggtcgggatc tcgtaagact acttcagaat gttgctagga
     1741 taccagaatt tgaactgctt tggaaagata ttatccataa tcctcaggcc ttgagtcctc
     1801 agttcacagg tatcctacag cttcttcagt caagaacatc ccgaaaattc ctagcatgtc
     1861 gtctaacccc ggacatggag actaaactcc tcttcatgac atcccgggtg cgatttggtc
     1921 aacaaaagcg ataccaagat tggttccagc gccagtacct gtcaactcca gatagtcagt
     1981 ctctgcgctg tgacctcatt cgctacatct gtggggtagt ccacccttct aatgaagtac
     2041 tgagttcaga tatcttgccc cggtgggcca tcattggttg gctcctgaca acgtgcacgt
     2101 caaatgtcgc tgcctccaat gccaagctgg ctttgtttta tgactggctg ttctttagtc
     2161 cagacaagga tagcattatg aacatagaac cagccatcct ggtcatgcac cactccatga
     2221 agccccaccc agccatcact gccacactcc tggacttcat gtgccgcatc attcccaact
     2281 tctatccacc attggagggc cacgtgcggc agggtgtctt ttcctccctc aaccacattg
     2341 tggagaaacg ggtcttggca cacctagctc ccctgtttga caaccctaag ttggataagg
     2401 agctgcgggc aatgctgaga gagaagtttc ctgagttctg cagctcaccc tccccacctg
     2461 tggaagtcaa aattgaggag ccagtttcca tggagatgga caaccatatg tcggataagg
     2521 atgagagttg ctatgacaat gcagaggcag ccttcagtga cgatgaagag gatctcaaca
     2581 gcaaaggaaa gaagagggag tttcgcttcc accctatcaa ggagacagtt gtggaggagc
     2641 cagttgatat caccccttac cttgaccagt tggatgagtc cctgagggac aaagtactcc
     2701 agctacagaa ggggagtgat acggaggccc agtgtgaggt catgcaggaa attgtggacc
     2761 aggtcctgga ggaagacttt gactcggagc agctgtctgt ccttgcttcc tgcctacagg
     2821 agctcttcaa ggcccacttt cgaggggagg tcctgcctga ggagattact gaggagtccc
     2881 tggaggagtc tgtaggaaag cctctgtacc taatatttag gaacctatgt cagatgcagg
     2941 aagacaacag cagcttctct ctacttctag accttctctc cgagctatat cagaagcagc
     3001 ccaagattgg ctaccacctg ctctactacc tgagggccag caaagccgcc gcagggaaga
     3061 tgaacctgta cgagtcattt gcccaggcta cccagctggg cgatctgcac acctgcctga
     3121 tgatggacat gaaggcctgc caggaggacg atgtgcggct cctgtgccac ctcacgccct
     3181 ccatctacac agagtttcca gatgaaacct tgaggagcgg agagctgctg aacatgatcg
     3241 tggctgttat tgactctgca cagctccagg agctggtctg ccacgtgatg atgggtaacc
     3301 tggttatgtt tcgaaaagac tcagttctca acatactcat tcagagccta gactgggaga
     3361 cctttgagca gtattgtgcc tggcagctct ttctggccca caatattccc ctggagacca
     3421 taatccccat cctgcagcac ctcaaataca aggagcaccc agaggccctg tcctgcctac
     3481 tgcttcaact ccgaagagaa aagcccagcg aggagatggt gaagatggtg ctgagccggc
     3541 cctgccatcc tgacgaccag ttcaccacca gcatcctgcg gcactggtgc atgaaacatg
     3601 acgagctgct ggccgagcac atcaagtccc tgctcatcaa gaacaacagc ctgcctcgca
     3661 agagacagag cctgaggagc tctagcagca agctggccca gctgactctg gagcagatcc
     3721 tggagcactt ggacaatctg cggctcaacc tgaccaacac caagcagaac ttttttagcc
     3781 agacgccaat tctccaggcg ctgcagcatg tccaagcgag ctgtgacgaa gcccacaaga
     3841 tgaaattcag tgatctcttc tccctggcgg aggaatatga ggactcttcc accaagccac
     3901 ccaagagccg gcgaaaagca gctctgtcca gccctcgaag tcgaaagaat gccacacagc
     3961 cccccaatgc cgaagaagag tcgggctcca gcagtgcttc agaagaggaa gacacgaaac
     4021 cgaagcctac caagcggaaa cgaaaagggt cctctgcagt gggctctgac agtgactgat
     4081 ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt
     4141 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4201 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     4261 atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg
     4321 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4381 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4441 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4501 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4561 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4621 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4681 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4741 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4801 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4861 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4921 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4981 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5041 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5101 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5161 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5221 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5281 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5341 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5401 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5461 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5521 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5581 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5641 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5701 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5761 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5821 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5881 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5941 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6001 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6061 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6121 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6181 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6241 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6301 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6361 aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     6421 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     6481 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     6541 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     6601 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     6661 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     6721 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     6781 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     6841 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     6901 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     6961 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     7021 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     7081 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     7141 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     7201 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     7261 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat
     7321 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     7381 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     7441 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7501 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7561 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7621 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg
     7681 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     7741 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     7801 agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc
     7861 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact
     7921 ggcagatgca cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca
     7981 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg
     8041 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaatt
//
