LOCUS       sequence-335147-        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RORg isoform 2 with N-terminal HisSUMO tag in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7232)
  AUTHORS   Strutzenberg TS, Zhu Y, Novick SJ, Garcia-Ordonez RD, Doebelin C, He
            Y, Chang MR, Kamenecka TM, Edwards DP, Griffin PR
  TITLE     Conformational Changes of RORgamma During Response Element 
            Recognition and Coregulator Engagement.
  JOURNAL   J Mol Biol. 2021 Sep 20;433(22):167258. doi: 
            10.1016/j.jmb.2021.167258.
  PUBMED    34547329
REFERENCE   2  (bases 1 to 7232)
  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: "J Mol 
            Biol."; date: "2021-09-20"; volume: "433(22):167258. doi"; pages: " 
            10.1016/j.jmb.2021.167258"
COMMENT     Sequence Label: pESUMO-hRORg2
FEATURES             Location/Qualifiers
     source          1..7232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             124..411
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             1964..1981
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2034..2052)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2048..2095
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2132..2587
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2219..2238)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2429..2450
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2613..2717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2718..2786,2787..3578)
                     /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(2936..2955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3749..4337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4238..4257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4491..4508
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4523..4662
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4588..4607)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4748..4770
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4764..4955)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5764..6846)
                     /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     6808..6827
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6847..6924)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7130..7149
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggtcatcacc atcatcatca cgggtccctg
      121 caggactcag aagtcaatca agaagctaag ccagaggtca agccagaagt caagcctgag
      181 actcacatca atttaaaggt gtccgatgga tcttcagaga tcttcttcaa gatcaaaaag
      241 accactcctt taagaaggct gatggaagcg ttcgctaaaa gacagggtaa ggaaatggac
      301 tccttaagat tcttgtacga cggtattaga attcaagctg atcaggcccc tgaagatttg
      361 gacatggagg ataacgatat tattgaggct caccgcgaac agattggagg tatgatgcgt
      421 acccagattg aagtgatccc gtgcaagatt tgcggtgata aaagcagcgg tatccactac
      481 ggcgttatta cctgcgaggg ttgcaagggc ttctttcgtc gtagccagcg ttgcaacgcg
      541 gcgtatagct gcacccgtca gcaaaactgc ccgatcgacc gtaccagccg taaccgttgc
      601 caacactgcc gtctgcagaa gtgcctggcg ctgggtatga gccgtgatgc ggtgaaattc
      661 ggccgtatga gcaagaaaca gcgtgacagc ctgcacgcgg aagtgcaaaa gcagctgcag
      721 caacgtcagc aacagcaaca ggaaccggtg gttaaaaccc cgccggcggg tgcgcagggt
      781 gcggacaccc tgacctacac cctgggtctg ccggatggtc aactgccgct gggtagcagc
      841 ccggacctgc cggaagcgag cgcgtgcccg ccgggcctgc tgaaggcgag cggtagcggc
      901 ccgagctata gcaacaacct ggcgaaagcg ggtctgaacg gtgcgagctg ccacctggag
      961 tacagcccgg aacgtggtaa agcggagggc cgtgaaagct tctatagcac cggtagccaa
     1021 ctgaccccgg atcgttgcgg cctgcgtttt gaggaacacc gtcatccggg tctgggtgaa
     1081 ctgggtcaag gtccggacag ctacggtagc ccgagcttcc gtagcacccc ggaagcgccg
     1141 tatgcgagcc tgaccgagat cgaacacctg gtgcagagcg tttgcaagag ctaccgtgaa
     1201 acctgccaac tgcgtctgga agatctgctg cgtcagcgta gcaacatttt tagccgtgag
     1261 gaagtgaccg gttaccaacg taaaagcatg tgggagatgt gggaacgttg cgcgcaccac
     1321 ctgaccgagg cgatccagta tgtggttgaa ttcgcgaagc gtctgagcgg ttttatggag
     1381 ctgtgccaaa acgatcagat tgtgctgctg aaagcgggcg cgatggaagt ggttctggtt
     1441 cgtatgtgcc gtgcgtacaa cgcggacaac cgtaccgttt tctttgaggg caagtatggt
     1501 ggcatggaac tgttccgtgc gctgggctgc agcgagctga tcagcagcat tttcgatttt
     1561 agccacagcc tgagcgcgct gcactttagc gaggacgaaa tcgcgctgta caccgcgctg
     1621 gtgctgatta acgcgcaccg tccgggtctg caagagaagc gtaaagttga acaactgcag
     1681 tataacctgg agctggcgtt tcaccaccac ctgtgcaaga cccaccgtca gagcattctg
     1741 gcgaaactgc cgccgaaggg caaactgcgt agcctgtgca gccaacacgt ggaacgtctg
     1801 caaatcttcc agcacctgca cccgattgtg gttcaggcgg cgtttccgcc gctgtataag
     1861 gaactgttta gcaccgaaac cgagagcccg gttggcctga gcaagtgata atctagagga
     1921 tccgaattcg agctccgtcg acaagcttgc ggccgcactc gagcaccacc accaccacca
     1981 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     2041 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     2101 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     2161 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2221 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2281 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2341 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2401 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2461 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2521 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2581 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2641 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2701 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2761 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2821 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2881 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2941 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3001 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3061 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3121 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3181 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3241 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3301 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3361 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3421 ccggtgagcg tggttctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3481 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3541 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3601 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3661 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3721 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3841 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3901 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3961 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4021 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4081 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4141 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4201 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4261 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4321 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4381 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4441 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4501 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4561 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4621 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4681 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4741 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4801 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4861 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4921 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4981 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     5041 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     5101 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     5161 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     5221 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5281 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     5341 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5401 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5461 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5521 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5581 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5641 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5701 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5761 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5821 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5881 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5941 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     6001 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     6061 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     6121 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     6181 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     6241 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     6301 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6361 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6421 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6481 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6541 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6601 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6661 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6721 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6781 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6841 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6901 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6961 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     7021 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     7081 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     7141 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     7201 cgtagaggat cgagatctcg atcccgcgaa at
//
