LOCUS       sequence-74334-e        7104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression vector for MBP-SRAP(13-236) fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  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 7104)
  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: hSRAP.HMG6.wt.Dlnk.a13-236
FEATURES             Location/Qualifiers
     source          1..7104
                     /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(853..873)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(901..1998)
                     /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(904..927)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     regulatory      1035..1044
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2005..2022)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2041..2063
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2078..2102
                     /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(2102..2121)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2103..2121)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2205..2224)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2430..2507
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2508..3590
                     /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(2527..2546)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4399..4590
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4584..4606)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4692..4834
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4747..4766
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4849..4866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5020..5608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5100..5119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5719..5738)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5730..6545
                     /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             5730..6545
                     /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(5803..5822)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6638..7093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6775..6796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6987..7006
                     /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 ggatgcttgt tggaaacccg gaatggtgtg atttttttct gccgtggctg ctgatttttc
      241 ttcgttggcc gcttcttcag aaaacaggct gcgtttttcg gcgatcagac gtttaacgcc
      301 caccatccac tgagaaactt ccgtcacgtg gtcgaccatc aggctgcgat gaatgtcatc
      361 agctgcatcc caacggtggg aactcagttc ctgaaccagc agtgccatac gttttttcac
      421 cgggatactc agtttaccgc cagcccattg ttcctgcagc agtgccaggc gacggctaat
      481 gtcatcacac acttgtttgc gcgtatgacc acggcagtct tccagagcct gttccagcgg
      541 acgcagcaca tcttccatga ctgcttccga ttcgaccggg aatgaggtcg gttcaacacc
      601 ggatgccgga ccactgccaa ccggcgggct acgcggtgct ttagagctcg gcggcggcgg
      661 acccatcggc ggcggacccg gcgaggtttc tgaggccggc acgcgcggag aaccatcttg
      721 cggtgctgcg acacgtttcg tcagcaggga gcgacgcgga ccgcccgctt gggtctgcag
      781 gccataacta aactgcggcg gatcattcca gccacgttct ttgttacccg gtttcacata
      841 cagttcagcc atgccctgga agtaaaggtt ttcggtcgtt gggatatcgt aatcggtacc
      901 agtctgcgcg tctttcaggg cttcatcgac agtctgacga ccgctggcgg cgttgatcac
      961 cgcagtacgc acggcatacc agaaagcgga catctgcggg atgttcggca tgatttcacc
     1021 tttctgggcg ttttccatgg tggcggcaat acgtggatct ttcgccaact cttcctcgta
     1081 agacttcagc gctacggcac ccagcggttt gtctttatta accgcttcca gaccttcatc
     1141 agtcagcaga tagttttcga ggaactcttt cgccagctct ttgttcggac tggcggcgtt
     1201 aatacctgcg ctcagcacgc caacgaacgg tttggatggt tgacccttga aggtcggcag
     1261 taccgttaca ccataattca ctttgctggt gtcgatgttg gaccatgccc acgggccgtt
     1321 gatggtcatc gctgtttcgc ctttattaaa ggcagcttct gcgatggagt aatcggtgtc
     1381 tgcattcatg tgtttgtttt taatcaggtc aaccaggaag gtcagacccg ctttcgcgcc
     1441 agcgttatcc acgcccacgt ctttaatgtc gtacttgccg ttttcatact tgaacgcata
     1501 acccccgtca gcagcaatca gcggccaggt gaagtacggt tcttgcaggt tgaacatcag
     1561 cgcgctctta cctttcgctt tcagttcttt atccagcgcc gggatctctt cccaggtttt
     1621 tggcgggttc ggcagcagat ctttgttata aatcagcgat aacgcttcaa cagcgatcgg
     1681 gtaagcaatc agcttgccgt tgtaacgtac ggcatcccag gtaaacggat acagcttgtc
     1741 ctggaacgct ttgtccgggg tgatttcagc caacaggcca gattgagcgt agccaccaaa
     1801 gcggtcgtgt gcccagaaga taatgtcagg gccatcgcca gttgccgcaa cctgtgggaa
     1861 tttctcttcc agtttatccg gatgctcaac ggtgacttta attccggtat ctttctcgaa
     1921 tttcttaccg acttcagcga gaccgttata gcctttatcg ccgttaatcc agattaccag
     1981 tttaccttct tcagttttgg taccgtgatg atgatgatga tggctgctgc ccatggtata
     2041 tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt
     2101 cccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg
     2161 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     2221 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     2281 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
     2341 ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
     2401 gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg
     2461 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     2521 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     2581 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     2641 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     2701 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     2761 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     2821 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     2881 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     2941 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     3001 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     3061 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     3121 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     3181 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     3241 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     3301 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     3361 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     3421 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     3481 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3541 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3601 attaatgtaa gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc
     3661 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     3721 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     3781 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     3841 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     3901 aagcaggcca ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg
     3961 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     4021 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     4081 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     4141 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     4201 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     4261 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     4321 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     4381 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     4441 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     4501 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     4561 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4621 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4681 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     4741 actatgcggc atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc
     4801 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     4861 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4921 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5461 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg
     5701 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     5761 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     5821 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     5881 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     5941 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     6001 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
     6061 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     6121 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     6181 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     6241 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     6301 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     6361 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     6421 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     6481 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     6541 tctaagaatt aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6601 ggggttccgc gcacatttcc ccgaaaagtg ccacctgaaa ttgtaaacgt taatattttg
     6661 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6721 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6781 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6841 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     6901 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6961 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7021 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7081 ctacagggcg cgtcccattc gcca
//
