LOCUS       sequence-315809-        7443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of NR2F2_Hormone-recep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  AUTHORS   Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz 
            AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
  TITLE     Large-scale survey and database of high affinity ligands for peptide
            recognition modules.
  JOURNAL   Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
  PUBMED    33438817
REFERENCE   2  (bases 1 to 7443)
  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: "Mol Syst 
            Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: " 
            10.15252/msb.20199310"
COMMENT     Sequence Label: pHH0103_NR2F2_Hormone-recep
FEATURES             Location/Qualifiers
     source          1..7443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(718..723,724..739,740..746)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    754..770
                     /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     759..781
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             855..872
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             888..1541
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1499..1521
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1548..1571
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1584..1601
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2355..2402
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3995..4450)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4132..4153)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4344..4363
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4492..4514)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4600..4740
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4655..4674
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4755..4772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4926..5513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5684..6475,6476..6544))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6307..6326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6545..6649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6717..6735
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6758..6835
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             join(6836..7443,1..475)
                     /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(6855..6874)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
       61 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
      121 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
      181 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
      241 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
      301 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
      361 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      421 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      481 acgcaattaa tgtaagttag ctcactcatt aggcacaatt ctcatgtttg acagcttatc
      541 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
      601 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
      661 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
      721 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
      781 gaaacagcca gtccgtttag gtgttttcac gagcacttca ccaacaagga ccatagatta
      841 tgaaaatcga agaacaccat caccatcacc attccagcgg taagcttatg tcccctatac
      901 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
      961 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
     1021 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
     1081 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
     1141 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
     1201 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
     1261 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     1321 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     1381 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     1441 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     1501 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcagatctg gaagttctgt
     1561 tccaggggcc cctgtccagc ggtctggttc cgcgtggttc cggtaccgcg gcccagccgg
     1621 cctcttacct gtctggctat atctctctgc tgctgcgtgc tgagccatac ccgacctccc
     1681 gtttcggctc tcagtgcatg cagccgaaca acatcatggg tattgaaaac atttgcgaac
     1741 tggcggcgcg tatgctgttt tccgccgttg aatgggctcg caacatcccg ttcttcccgg
     1801 acctgcagat tactgatcag gtggcactgc tgcgtctgac ctggtccgaa ctgttcgtgc
     1861 tgaatgcagc acaatgttct atgccactgc atgtagcgcc gctgctggct gcagccggtc
     1921 tgcacgctag cccgatgagc gcggaccgtg ttgttgcgtt tatggaccac atccgcatct
     1981 tccaggagca ggtggagaaa ctgaaagcgc tgcacgtgga tagcgctgag tactcttgcc
     2041 tgaaggccat cgtcctgttt acctctgatg cgtgcggcct gagcgatgtt gctcacgttg
     2101 aaagcctgca ggaaaaaagc cagtgtgcac tggaagaata cgttcgttct cagtacccta
     2161 accagccgac tcgtttcggt aaactgctgc tgcgcctgcc gtccctgcgt accgtctctt
     2221 cctctgttat cgaacaactg ttcttcgtac gcctggtagg taaaaccccg attgaaacgc
     2281 tgatccgtga catgctgctg tccggttcca gcttcaactg gccgtatatg gccatccagg
     2341 cggccgcata ataaaccgat acaattaaag gctccttttg gagccttttt tttggagatt
     2401 ttcaacgtga aaaaattatt attcgcaatt cctttagttg ttcctttcta ttctcactcc
     2461 gctgaaactg ttgaaagttg tttagcaaaa ccccatacag aaaattcatt tactaacgtc
     2521 tggaaagacg acaaaacttt agatcgttac gctaactatg agggctgtct gtggaatgct
     2581 acaggcgttg tagtttgtac tggtgacgaa actcagtgtc tagctagagt ggcggtggct
     2641 ctggttccgg tgattttgat tatgaaaaga tggcaaacgc taataagggg gctatgaccg
     2701 aaaatgccga tgaaaacgcg ctacagtctg acgctaaagg caaacttgat tctgtcgcta
     2761 ctgattacgg tgctgctatc gatggtttca ttggtgacgt ttccggcctt gctaatggta
     2821 atggtgctac tggtgatttt gctggctcta attcccaaat ggctcaagtc ggtgacggtg
     2881 ataattcacc tttaatgaat aatttccgtc aatatttacc ttccctccct caatcggttg
     2941 aatgtcgccc ttttgtcttt ggcgctggta aaccatatga attttctatt gattgtgaca
     3001 aaataaactt attccgtggt gtctttgcgt ttcttttata tgttgccacc tttatgtatg
     3061 tattttctac gtttgctaac atactgcgta ataaggagtc ttaatcatgc cagttctttt
     3121 ggctagcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     3181 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
     3241 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     3301 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
     3361 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     3421 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     3481 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     3541 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     3601 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     3661 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     3721 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     3781 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     3841 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     3901 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     3961 gaccacgctg atgagcttta ccgcaggatc cgcgaaattg taagcgttaa tattttgtta
     4021 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4081 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4141 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4201 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4261 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4321 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4381 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4441 cagggcgcgt cgcggatcct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4501 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4561 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     4621 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4681 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4741 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4801 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4861 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4981 aggtggcgaa acccgacagg actataaaga taccagggtt tccccctgga agctccctcg
     5041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5101 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5341 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5461 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5521 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5581 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5641 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5701 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5761 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5821 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5881 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5941 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     6001 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6061 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6121 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6181 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6241 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6301 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6361 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6421 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6481 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6541 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6601 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6661 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     6721 ggcgtatcac gaggcccttt cgtcttcaag aattcccgac accatcgaat ggtgcaaaac
     6781 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     6841 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     6901 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     6961 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     7021 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     7081 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     7141 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     7201 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     7261 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     7321 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     7381 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     7441 taa
//
