LOCUS       sequence-74343-e        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression vector for MBP-SLIRP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6786)
  AUTHORS   McKay DB, Xi L, Barthel KK, Cech TR
  TITLE     Structure and function of steroid receptor RNA activator protein, 
            the proposed partner of SRA noncoding RNA.
  JOURNAL   J Mol Biol. 2014 Apr 17;426(8):1766-85. doi: 
            10.1016/j.jmb.2014.01.006. Epub 2014 Jan 30.
  PUBMED    24486609
REFERENCE   2  (bases 1 to 6786)
  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: 
            "10.1016/j.jmb.2014.01.006"; journalName: "J Mol Biol"; date: 
            "2014-04-17- 17"; volume: "426"; issue: "8"; pages: "1766-85"
COMMENT     Sequence Label: SLIRP.HMG6.wt.fl
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(535..555)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(583..1680)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(586..609)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     regulatory      717..726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1687..1704)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1723..1745
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1760..1784
                     /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     complement(1784..1803)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1785..1803)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1887..1906)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2112..2189
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2190..3272
                     /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(2209..2228)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4081..4272
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4266..4288)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4374..4516
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4429..4448
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4531..4548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4702..5290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4782..4801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5401..5420)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5412..6227
                     /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             5412..6227
                     /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     complement(5485..5504)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6320..6775)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6457..6478)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6669..6688
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gaggcggccg cggtacccta
      181 gaagtctttt ttttcatcat cgctggtttg cggcagtttc ggacgacggg tgtggacttg
      241 aactttcaca ccgtcgataa tgtgattttc ttgctgcagc gcgttgcgca ggccttcttc
      301 gctactaaac tgaacccaac ccaggccacg atggaaaccg gtttctttat caaacggcag
      361 aatgcagcga cggacgtggc cgaattgtgc aaaatgttct ttcagctgag agctggccgc
      421 ggtccacggg atacgacgaa cgaacgccac cggttgattg atcgagcgac gcagggcagc
      481 agcaccacgg gcagccgacg cagccatatg gccgccgccg ccgccgccgc tatggccctg
      541 gaagtaaagg ttttcggtcg ttgggatatc gtaatcggta ccagtctgcg cgtctttcag
      601 ggcttcatcg acagtctgac gaccgctggc ggcgttgatc accgcagtac gcacggcata
      661 ccagaaagcg gacatctgcg ggatgttcgg catgatttca cctttctggg cgttttccat
      721 ggtggcggca atacgtggat ctttcgccaa ctcttcctcg taagacttca gcgctacggc
      781 acccagcggt ttgtctttat taaccgcttc cagaccttca tcagtcagca gatagttttc
      841 gaggaactct ttcgccagct ctttgttcgg actggcggcg ttaatacctg cgctcagcac
      901 gccaacgaac ggtttggatg gttgaccctt gaaggtcggc agtaccgtta caccataatt
      961 cactttgctg gtgtcgatgt tggaccatgc ccacgggccg ttgatggtca tcgctgtttc
     1021 gcctttatta aaggcagctt ctgcgatgga gtaatcggtg tctgcattca tgtgtttgtt
     1081 tttaatcagg tcaaccagga aggtcagacc cgctttcgcg ccagcgttat ccacgcccac
     1141 gtctttaatg tcgtacttgc cgttttcata cttgaacgca taacccccgt cagcagcaat
     1201 cagcggccag gtgaagtacg gttcttgcag gttgaacatc agcgcgctct tacctttcgc
     1261 tttcagttct ttatccagcg ccgggatctc ttcccaggtt tttggcgggt tcggcagcag
     1321 atctttgtta taaatcagcg ataacgcttc aacagcgatc gggtaagcaa tcagcttgcc
     1381 gttgtaacgt acggcatccc aggtaaacgg atacagcttg tcctggaacg ctttgtccgg
     1441 ggtgatttca gccaacaggc cagattgagc gtagccacca aagcggtcgt gtgcccagaa
     1501 gataatgtca gggccatcgc cagttgccgc aacctgtggg aatttctctt ccagtttatc
     1561 cggatgctca acggtgactt taattccggt atctttctcg aatttcttac cgacttcagc
     1621 gagaccgtta tagcctttat cgccgttaat ccagattacc agtttacctt cttcagtttt
     1681 ggtaccgtga tgatgatgat gatggctgct gcccatggta tatctccttc ttaaagttaa
     1741 acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta
     1801 ttaatttcgc gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac
     1861 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     1921 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     1981 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     2041 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     2101 tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     2161 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
     2221 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     2281 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     2341 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     2401 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     2461 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     2521 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     2581 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     2641 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     2701 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     2761 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     2821 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     2881 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     2941 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     3001 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
     3061 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     3121 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
     3181 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3241 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc
     3301 actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     3361 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     3421 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     3481 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     3541 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     3601 ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc
     3661 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
     3721 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
     3781 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc
     3841 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg
     3901 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc
     3961 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc
     4021 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca
     4081 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
     4141 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
     4201 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
     4261 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4321 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     4381 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     4441 agattgtact gagagtgcac catatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4501 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5101 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5281 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5341 ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata aacagtaata
     5401 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg cgattaaatt
     5461 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     5521 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     5581 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     5641 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     5701 tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     5761 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     5821 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     5881 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     5941 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     6001 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     6061 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     6121 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     6181 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa ttaattcatg
     6241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6301 ccccgaaaag tgccacctga aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     6361 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6421 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6481 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6541 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6601 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6661 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6721 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     6781 tcgcca
//
