LOCUS       sequence-246093-        6505 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 6505)
  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 6505)
  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..6505
                     /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(2555..2577)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2722..2740)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2808..2912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2913..2981,2982..3773)
                     /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(3131..3150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3944..4532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4433..4452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4686..4703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4776..4853
                     /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             4854..5936
                     /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(4873..4892)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5949..5970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5985..5990,5991..6008,6009..6015)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6023..6039
                     /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     6047..6063
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6047..6063
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6059..6505,1..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(6079..6096)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6079..6095)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6088..6110)
                     /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 ccagtttccc tcgagcggcc gcatcgtgac tgactgacga
     2521 tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2581 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2641 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     2701 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2761 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2821 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2881 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2941 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3001 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3061 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3121 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     3181 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3241 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3301 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3361 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3421 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     3481 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3541 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3601 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3661 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3721 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3781 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3841 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3961 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4021 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4081 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4141 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4201 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4261 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4321 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4381 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4441 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4501 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4561 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4621 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4681 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4741 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg tgcaaaacct
     4801 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     4861 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     4921 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     4981 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     5041 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     5101 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5161 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5221 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     5281 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     5341 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     5401 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     5461 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     5521 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     5581 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     5641 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     5701 tcccgccgtc aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     5761 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     5821 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5881 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5941 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6001 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6061 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6121 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6181 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
     6241 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
     6301 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
     6361 caccaacgta acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
     6421 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
     6481 aattattttt gatggcgttg gaatt
//
