LOCUS       sequence-211959-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes C-terminal part of RANKL (aa158-aa316) with a GST N-terminal
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  AUTHORS   Beranger GE, Momier D, Guigonis JM, Samson M, Carle GF, Scimeca JC
  TITLE     Differential binding of poly(ADP-Ribose) polymerase-1 and JunD/Fra2 
            accounts for RANKL-induced Tcirg1 gene expression during 
            osteoclastogenesis.
  JOURNAL   J Bone Miner Res. 2007 Jul;22(7):975-83. doi: 10.1359/jbmr.070406.
  PUBMED    17419679
REFERENCE   2  (bases 1 to 5447)
  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: "10.1359/jbmr"; 
            journalName: "J Bone Miner Res"; date: "2007-07"; volume: "22"; 
            issue: "7"; pages: "975-83"
COMMENT     Sequence Label: pGEX4T2 RANK-L
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(137..142,143..160,161..167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    175..191
                     /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     199..215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     199..215
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             211..738
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(231..248)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(231..247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(240..262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(840..845,846..861,862..868)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    876..892
                     /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)."
     CDS             915..1568
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1526..1548
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1575..1592
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2154..2176)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2321..2339)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2407..2511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2512..2580,2581..3372)
                     /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     complement(2730..2749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3543..4131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4032..4051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4285..4302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4375..4452
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4453..5447,1..88)
                     /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(4472..4491)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
       61 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
      121 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      181 gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc
      241 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      301 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      361 cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg
      421 ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac
      481 tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc cattacggtc
      541 aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt
      601 gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt tggaattagc
      661 ttatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta
      721 tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct
      781 ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt
      841 tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac
      901 aggaaacagt attcatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac
      961 ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg
     1021 atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc
     1081 cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag
     1141 ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg
     1201 aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg
     1261 aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc
     1321 gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt
     1381 atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat
     1441 tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca
     1501 gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc
     1561 ctccaaaatc ggatctggtt ccgcgtggat ccccaggaat tcccgggtcg acaaagcctg
     1621 aggcccagcc atttgcacac ctcaccatca atgctgccag catcccatcg ggttcccata
     1681 aagtcactct gtcctcttgg taccacgatc gaggctgggc caagatctct aacatgacgt
     1741 taagcaacgg aaaactaagg gttaaccaag atggcttcta ttacctgtac gccaacattt
     1801 gctttcggca tcatgaaaca tcgggaagcg tacctacaga ctatcttcag ctgatggtgt
     1861 atgtcgttaa aaccagcatc aaaatcccaa gttctcataa cctgatgaaa ggagggagca
     1921 cgaaaaactg gtcgggcaat tctgaattcc acttttattc cataaatgtt gggggatttt
     1981 tcaagctccg agctggtgaa gaaattagca ttcaggtgtc caacccttcc ctgctggatc
     2041 cggatcaaga tgcgacgtac tttggggctt tcaaagttca ggacatagac tgagcggccg
     2101 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     2161 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2221 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     2281 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     2341 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2401 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2461 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2521 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2581 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2641 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2701 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2761 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2821 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2881 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2941 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3001 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     3061 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3121 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3181 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3241 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3301 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3361 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3421 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3481 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3541 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3601 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3661 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3721 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3781 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3841 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3901 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3961 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4021 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4081 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4141 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4201 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4261 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4321 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
     4381 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     4441 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     4501 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     4561 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     4621 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     4681 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     4741 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     4801 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     4861 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     4921 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     4981 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     5041 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     5101 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     5161 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     5221 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     5281 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     5341 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     5401 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcc
//
