LOCUS       sequence-252201-        5550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express C9ORF80 in E. coli with a His tag (C-terminal).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  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 5550)
  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: pET28a-C9ORF80
FEATURES             Location/Qualifiers
     source          1..5550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     219..238
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        219..237
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    238..262
                     /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)."
     RBS             277..299
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             633..650
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(703..721)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      717..764
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      801..1256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(888..907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1098..1119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1348..2163)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1348..2163)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2071..2090
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2155..2174
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2285..2873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2774..2793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3027..3044
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3059..3198
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3124..3143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3284..3306
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3300..3491)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4300..5382)
                     /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     5344..5363
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5383..5460)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
       61 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
      121 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
      181 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga
      241 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      301 ataccatggc agcaaactct tcaggacaag gttttcaaaa caaaaataga gttgcaatct
      361 tggcagaact ggacaaagag aaaagaaaac tacttatgca gaaccagtct tcaacaaatc
      421 atcctggagc tagcattgca ctctcgagac cctctcttaa taaggacttc cgggatcacg
      481 ctgagcagca gcatattgca gcccaacaga aggcagcttt gcagcatgct catgcacatt
      541 catctggata cttcatcact caagactctg catttgggaa ccttattctt cctgttttac
      601 ctcgccttga cccagaagcg gccgcactcg agcaccacca ccaccaccac tgagatccgg
      661 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
      721 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
      781 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
      841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
      901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
      961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1141 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1201 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     1261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1321 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
     1381 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     1441 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     1501 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     1561 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     1621 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     1681 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     1741 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     1801 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     1861 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     1921 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     1981 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     2041 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     2101 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     2161 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa
     2221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2401 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2941 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3001 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3061 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     3121 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3181 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3241 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3301 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3361 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3421 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     3481 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     3541 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     3601 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     3661 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     3721 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     3781 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     3841 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     3901 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg
     3961 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4021 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4081 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     4141 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     4201 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     4261 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4321 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4381 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     4441 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     4501 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     4561 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     4621 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     4681 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     4741 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     4801 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     4861 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     4921 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     4981 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5041 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     5101 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     5161 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     5221 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     5281 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     5341 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     5401 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     5461 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     5521 tgaggccgtt gagcaccgcc gccgcaagga
//
