LOCUS       sequence-350019-        6325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of TPR2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6325)
  AUTHORS   Ke J, Ma H, Gu X, Thelen A, Brunzelle JS, Li J, Xu HE, Melcher K
  TITLE     Structural basis for recognition of diverse transcriptional 
            repressors by the TOPLESS family of corepressors.
  JOURNAL   Sci Adv. 2015 Jul 24;1(6):e1500107. doi: 10.1126/sciadv.1500107. 
            eCollection 2015 Jul.
  PUBMED    26601214
REFERENCE   2  (bases 1 to 6325)
  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"; journalName: "Sci Adv."; 
            date: "2015-07-24"; volume: "1(6):e1500107. doi"; pages: " 
            10.1126/sciadv.1500107. eCollection 2015 Jul"
COMMENT     Sequence Label: pSumo-His6-SUMO-AtTPR2(1-209)
FEATURES             Location/Qualifiers
     source          1..6325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..78)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     284..303
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     387..406
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        387..405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    406..430
                     /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             445..467
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             480..497
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             501..791
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             1443..1460
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1513..1531)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1527..1574
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1611..2066
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1698..1717)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1908..1929
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2092..2196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2197..2265,2266..3057)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2415..2434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3228..3816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3717..3736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3970..3987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4002..4141
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4067..4086)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4227..4249
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4243..4434)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5243..6325)
                     /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     6287..6306
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
       61 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
      121 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
      181 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
      241 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      301 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      361 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
      421 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggtc
      481 atcaccatca tcatcacggg tcggactcag aagtcaatca agaagctaag ccagaggtca
      541 agccagaagt caagcctgag actcacatca atttaaaggt gtccgatgga tcttcagaga
      601 tcttcttcaa gatcaaaaag accactcctt taagaaggct gatggaagcg ttcgctaaaa
      661 gacagggtaa ggaaatggac tccttaagat tcttgtacga cggtattaga attcaagctg
      721 atcaggcccc tgaagatttg gacatggagg ataacgatat tattgaggct caccgcgaac
      781 agattggagg tggatccatg agcagtctga gtcgcgagct ggtgtttctg atcctgcaat
      841 tcctggacga ggagaagttc aaggagagcg tgcacaagct ggagcaggag agcggcttct
      901 tcttcaacat taagtacttc gaagagaaag cactggccgg cgaatgggac gaggttgaga
      961 agtacctgag cggctttacc aaggtggacg acaaccgcta cagcatgaaa attttcttcg
     1021 agatccgcaa acaaaagtac ctggaggccc tggaccgcaa cgatcgtgcc aaggccgtgg
     1081 agatcctggc caaggacctg aaggtgttcg ccaccttcaa cgaggagctg tacaaggaga
     1141 tcacccagct gctgacactg gagaacttcc gcgagaacga acagctgagc aagtacggcg
     1201 acaccaaaag cgcacgcagc atcatgtaca ccgagctgaa gaagctgatc gaggccaacc
     1261 cgctgttccg cgagaagctg gccttcccga gcttcaaggc cagtcgcctg cgcaccctga
     1321 tcaaccaaag cctgaactgg cagcaccagc tgtgcaagaa cccgcgtccg aacccggaca
     1381 ttaagaccct gttcctggac cacagttgta gtccgagcaa tggctaagcg gccgcactcg
     1441 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     1501 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1561 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     1621 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1681 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1741 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1801 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1861 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1921 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1981 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2041 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     2101 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2161 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2401 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2581 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2701 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     2761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2881 attgctgata aatctggagc cggtgagcgt ggttctcgcg gtatcattgc agcactgggg
     2941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3061 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3121 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3181 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3241 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3301 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3361 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3421 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3481 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3541 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3601 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3661 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3721 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3781 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3841 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3901 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3961 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4021 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     4081 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     4141 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4201 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4261 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     4321 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     4381 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     4441 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     4501 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     4561 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     4621 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     4681 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     4741 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     4801 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     4861 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat
     4921 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     4981 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     5041 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     5101 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     5161 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     5221 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5281 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     5341 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     5401 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     5461 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     5521 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     5581 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     5641 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     5701 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     5761 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     5821 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     5881 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     5941 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     6001 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     6061 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     6121 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     6181 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     6241 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     6301 catcgtataa cgttactggt ttcac
//
