LOCUS       sequence-252287-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express INTS3 C-termini (514-1042 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 6557)
  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 6557)
  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-C
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(148..153,154..169,170..176)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    184..200
                     /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             223..876
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     834..856
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             883..906
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2572..2594)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2739..2757)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2825..2929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2930..2998,2999..3790)
                     /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(3148..3167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3961..4549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4450..4469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4703..4720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4793..4870
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4871..5953
                     /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(4890..4909)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6002..6007,6008..6025,6026..6032)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6040..6056
                     /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     6064..6080
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6064..6080
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6076..6557,1..46)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6096..6113)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6096..6112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6105..6127)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt
       61 cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg
      121 ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg
      181 gaattgtgag cggataacaa tttcacacag gaaacagtat tcatgtcccc tatactaggt
      241 tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa
      301 aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt
      361 gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca
      421 cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca
      481 aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt
      541 tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta
      601 cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat
      661 catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac
      721 ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc
      781 ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg
      841 caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag
      901 gggcccctgg gatccatgga gatggacaac catatgtcgg ataaggatga gagttgctat
      961 gacaatgcag aggcagcctt cagtgacgat gaagaggatc tcaacagcaa aggaaagaag
     1021 agggagtttc gcttccaccc tatcaaggag acagttgtgg aggagccagt tgatatcacc
     1081 ccttaccttg accagttgga tgagtccctg agggacaaag tactccagct acagaagggg
     1141 agtgatacgg aggcccagtg tgaggtcatg caggaaattg tggaccaggt cctggaggaa
     1201 gactttgact cggagcagct gtctgtcctt gcttcctgcc tacaggagct cttcaaggcc
     1261 cactttcgag gggaggtcct gcctgaggag attactgagg agtccctgga ggagtctgta
     1321 ggaaagcctc tgtacctaat atttaggaac ctatgtcaga tgcaggaaga caacagcagc
     1381 ttctctctac ttctagacct tctctccgag ctatatcaga agcagcccaa gattggctac
     1441 cacctgctct actacctgag ggccagcaaa gccgccgcag ggaagatgaa cctgtacgag
     1501 tcatttgccc aggctaccca gctgggcgat ctgcacacct gcctgatgat ggacatgaag
     1561 gcctgccagg aggacgatgt gcggctcctg tgccacctca cgccctccat ctacacagag
     1621 tttccagatg aaaccttgag gagcggagag ctgctgaaca tgatcgtggc tgttattgac
     1681 tctgcacagc tccaggagct ggtctgccac gtgatgatgg gtaacctggt tatgtttcga
     1741 aaagactcag ttctcaacat actcattcag agcctagact gggagacctt tgagcagtat
     1801 tgtgcctggc agctctttct ggcccacaat attcccctgg agaccataat ccccatcctg
     1861 cagcacctca aatacaagga gcacccagag gccctgtcct gcctactgct tcaactccga
     1921 agagaaaagc ccagcgagga gatggtgaag atggtgctga gccggccctg ccatcctgac
     1981 gaccagttca ccaccagcat cctgcggcac tggtgcatga aacatgacga gctgctggcc
     2041 gagcacatca agtccctgct catcaagaac aacagcctgc ctcgcaagag acagagcctg
     2101 aggagctcta gcagcaagct ggcccagctg actctggagc agatcctgga gcacttggac
     2161 aatctgcggc tcaacctgac caacaccaag cagaactttt ttagccagac gccaattctc
     2221 caggcgctgc agcatgtcca agcgagctgt gacgaagccc acaagatgaa attcagtgat
     2281 ctcttctccc tggcggagga atatgaggac tcttccacca agccacccaa gagccggcga
     2341 aaagcagctc tgtccagccc tcgaagtcga aagaatgcca cacagccccc caatgccgaa
     2401 gaagagtcgg gctccagcag tgcttcagaa gaggaagaca cgaaaccgaa gcctaccaag
     2461 cggaaacgaa aagggtcctc tgcagtgggc tctgacagtg actgatctcg agcggccgca
     2521 tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2581 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2641 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     2701 tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt
     2761 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     2821 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2941 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3001 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3061 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3121 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     3181 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3241 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3301 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3361 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3421 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     3481 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3541 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3601 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3661 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3721 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3781 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3841 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3901 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3961 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4021 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4081 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4141 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4201 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4261 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4321 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4381 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4441 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4501 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4561 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4621 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4681 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     4741 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat
     4801 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     4861 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     4921 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     4981 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     5041 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     5101 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     5161 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     5221 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     5281 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     5341 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     5401 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     5461 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     5521 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     5581 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     5641 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     5701 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     5761 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     5821 gctgttgccc gtctcactgg tgaaaagaaa aaccacgcta gcgcccaata cgcaaaccgc
     5881 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5941 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg
     6001 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     6061 acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt
     6121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     6181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     6241 aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg
     6301 gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt
     6361 tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt
     6421 cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta
     6481 caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg
     6541 tgcaccaatg cttctgg
//
