LOCUS       sequence-74331-e        7140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression vector for MBP-SRAP(1-236) fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7140)
  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 7140)
  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.Dlink.a1-236
FEATURES             Location/Qualifiers
     source          1..7140
                     /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(889..909)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(937..2034)
                     /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(940..963)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     regulatory      1071..1080
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2041..2058)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2077..2099
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2114..2138
                     /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(2138..2157)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2139..2157)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2241..2260)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2466..2543
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2544..3626
                     /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(2563..2582)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4435..4626
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4620..4642)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4728..4870
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4783..4802
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4885..4902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5056..5644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5136..5155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5755..5774)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5766..6581
                     /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             5766..6581
                     /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(5839..5858)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6674..7129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6811..6832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7023..7042
                     /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 atttccactt ctgcttgacc tgccggacag cgcgtcatgc cctggaagta
      901 aaggttttcg gtcgttggga tatcgtaatc ggtaccagtc tgcgcgtctt tcagggcttc
      961 atcgacagtc tgacgaccgc tggcggcgtt gatcaccgca gtacgcacgg cataccagaa
     1021 agcggacatc tgcgggatgt tcggcatgat ttcacctttc tgggcgtttt ccatggtggc
     1081 ggcaatacgt ggatctttcg ccaactcttc ctcgtaagac ttcagcgcta cggcacccag
     1141 cggtttgtct ttattaaccg cttccagacc ttcatcagtc agcagatagt tttcgaggaa
     1201 ctctttcgcc agctctttgt tcggactggc ggcgttaata cctgcgctca gcacgccaac
     1261 gaacggtttg gatggttgac ccttgaaggt cggcagtacc gttacaccat aattcacttt
     1321 gctggtgtcg atgttggacc atgcccacgg gccgttgatg gtcatcgctg tttcgccttt
     1381 attaaaggca gcttctgcga tggagtaatc ggtgtctgca ttcatgtgtt tgtttttaat
     1441 caggtcaacc aggaaggtca gacccgcttt cgcgccagcg ttatccacgc ccacgtcttt
     1501 aatgtcgtac ttgccgtttt catacttgaa cgcataaccc ccgtcagcag caatcagcgg
     1561 ccaggtgaag tacggttctt gcaggttgaa catcagcgcg ctcttacctt tcgctttcag
     1621 ttctttatcc agcgccggga tctcttccca ggtttttggc gggttcggca gcagatcttt
     1681 gttataaatc agcgataacg cttcaacagc gatcgggtaa gcaatcagct tgccgttgta
     1741 acgtacggca tcccaggtaa acggatacag cttgtcctgg aacgctttgt ccggggtgat
     1801 ttcagccaac aggccagatt gagcgtagcc accaaagcgg tcgtgtgccc agaagataat
     1861 gtcagggcca tcgccagttg ccgcaacctg tgggaatttc tcttccagtt tatccggatg
     1921 ctcaacggtg actttaattc cggtatcttt ctcgaatttc ttaccgactt cagcgagacc
     1981 gttatagcct ttatcgccgt taatccagat taccagttta ccttcttcag ttttggtacc
     2041 gtgatgatga tgatgatggc tgctgcccat ggtatatctc cttcttaaag ttaaacaaaa
     2101 ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt
     2161 tcgcgggatc gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     2221 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     2281 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     2341 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     2401 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga
     2461 tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     2521 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     2581 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     2641 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     2701 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     2761 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     2821 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     2881 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     2941 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     3001 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     3061 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     3121 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     3181 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     3241 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     3301 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     3361 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     3421 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     3481 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     3541 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3601 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     3661 taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     3721 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     3781 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     3841 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     3901 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     3961 gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     4021 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     4081 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     4141 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     4201 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     4261 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     4321 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     4381 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     4441 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     4501 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     4561 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     4621 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4681 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4741 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4801 tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4861 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5461 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5701 ttaagggatt ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     5761 gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca
     5821 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     5881 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     5941 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     6001 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     6061 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     6121 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     6181 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     6241 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     6301 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     6361 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     6421 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     6481 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     6541 ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt catgagcgga
     6601 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6661 aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     6721 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6781 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6841 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6901 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6961 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     7021 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     7081 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
