LOCUS       sequence-252290-        6508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express INTS3 N-termini (1-513 aa) 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 6508)
  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 6508)
  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-N
FEATURES             Location/Qualifiers
     source          1..6508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(673..678,679..696,697..703)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    711..727
                     /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     735..751
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     735..751
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             747..1274
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(767..784)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(767..783)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(776..798)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1376..1381,1382..1397,1398..1404)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1412..1428
                     /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             1451..2104
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2062..2084
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2111..2134
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3751..3773)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3918..3936)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4004..4108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4109..4177,4178..4969)
                     /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(4327..4346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5140..5728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5629..5648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5882..5899
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5972..6049
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6050..6508,1..624)
                     /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(6069..6088)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
       61 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
      121 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
      181 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
      241 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
      301 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
      361 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
      421 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
      481 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccacgct agcgcccaat
      541 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      601 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
      661 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      721 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt
      781 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      841 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      901 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
      961 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
     1021 agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc
     1081 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
     1141 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga attagcttat
     1201 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     1261 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     1321 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     1381 aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
     1441 aacagtattc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     1501 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     1561 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     1621 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     1681 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     1741 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     1801 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     1861 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     1921 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     1981 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     2041 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     2101 aaaatcggat ctggaagttc tgttccaggg gcccctggga tccatggagt tgcagaaggg
     2161 aaaaggggcg gcagcagcag cagctgcttc gggagcagcg ggaggtggag gaggaggagc
     2221 gggagcagga gccccaggag gggggaggct gctactttca accagtttgg atgccaagga
     2281 tgagttagag gagagattgg aaaggtgtat gagcattgtg acatcgatga ctgctggtgt
     2341 ctcggagaga gaagccaatg atgccctcaa tgcgtatgtg tgcaaaggcc tcccccagca
     2401 tgaagaaatc tgcctgggcc tgtttactct catcctcact gaacctgccc aagcccagaa
     2461 gtgttaccgg gacttagctc tggtgagtcg tgatggcatg aatattgtcc tgaataaaat
     2521 caaccagata cttatggaga agtacctgaa gctgcaggat acctgccgta ctcagttggt
     2581 gtggttggta cgggaactgg tgaagagtgg ggttctggga gccgatggtg tttgtatgac
     2641 gtttatgaag cagattgcag gtggagatgt tacagccaaa aatatctggt tggcagaaag
     2701 tgttctggat atcctgacag agcaaaggga gtgggtcctg aagagcagca tcctcattgc
     2761 catggctgtt tacacgtacc tccgcctcat cgtggaccac catgggactg cccagctcca
     2821 ggccctgcga cagaaggaag tagacttctg catctcactg cttcgggaac ggttcatgga
     2881 atgtctgatg attggtcggg atctcgtaag actacttcag aatgttgcta ggataccaga
     2941 atttgaactg ctttggaaag atattatcca taatcctcag gccttgagtc ctcagttcac
     3001 aggtatccta cagcttcttc agtcaagaac atcccgaaaa ttcctagcat gtcgtctaac
     3061 cccggacatg gagactaaac tcctcttcat gacatcccgg gtgcgatttg gtcaacaaaa
     3121 gcgataccaa gattggttcc agcgccagta cctgtcaact ccagatagtc agtctctgcg
     3181 ctgtgacctc attcgctaca tctgtggggt agtccaccct tctaatgaag tactgagttc
     3241 agatatcttg ccccggtggg ccatcattgg ttggctcctg acaacgtgca cgtcaaatgt
     3301 cgctgcctcc aatgccaagc tggctttgtt ttatgactgg ctgttcttta gtccagacaa
     3361 ggatagcatt atgaacatag aaccagccat cctggtcatg caccactcca tgaagcccca
     3421 cccagccatc actgccacac tcctggactt catgtgccgc atcattccca acttctatcc
     3481 accattggag ggccacgtgc ggcagggtgt cttttcctcc ctcaaccaca ttgtggagaa
     3541 acgggtcttg gcacacctag ctcccctgtt tgacaaccct aagttggata aggagctgcg
     3601 ggcaatgctg agagagaagt ttcctgagtt ctgcagctca ccctccccac ctgtggaagt
     3661 caaaattgag gagccagttt cctgactcga gcggccgcat cgtgactgac tgacgatctg
     3721 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3781 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3841 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa
     3901 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3961 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4021 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4081 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4141 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4201 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4261 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4321 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4381 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4441 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4501 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4561 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4621 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     4681 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4741 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4801 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4861 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4921 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4981 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5041 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5101 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5161 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5221 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5281 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5341 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5401 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5461 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5521 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5581 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5641 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5701 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5761 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5821 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5881 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     5941 tctgtgcggt atttcacacc gcataaattc cgacaccatc gaatggcgca aaacctttcg
     6001 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     6061 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     6121 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     6181 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     6241 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     6301 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     6361 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     6421 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     6481 ggcgttattt cttgatgtct ctgaccag
//
