LOCUS       sequence-259627-        6941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mouse Tankyrase1 ARC2-3 (308-655) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6941)
  AUTHORS   Morrone S, Cheng Z, Moon RT, Cong F, Xu W
  TITLE     Crystal structure of a Tankyrase-Axin complex and its implications 
            for Axin turnover and Tankyrase substrate recruitment.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Jan 31;109(5):1500-5. doi: 
            10.1073/pnas.1116618109. Epub 2012 Jan 17.
  PUBMED    22307604
REFERENCE   2  (bases 1 to 6941)
  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: 
            "10.1073/pnas.1116618109"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-01-31- 31"; volume: "109"; issue: "5"; pages: "1500-5"
COMMENT     Sequence Label: pGEX-4T1 mouse Tankyrase1 ARC2-3 (308-655)
FEATURES             Location/Qualifiers
     source          1..6941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..717
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             727..747
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2773..2795)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2940..2958)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3026..3130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3131..3199,3200..3991)
                     /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(3349..3368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4162..4750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4651..4670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4904..4921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4994..5071
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5072..6154
                     /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(5091..5110)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6203..6208,6209..6226,6227..6233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6241..6257
                     /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     6265..6281
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6265..6281
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6277..6804
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6297..6314)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6297..6313)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6306..6328)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6906..6911,6912..6927,6928..6934)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggttccgcg tggatccccg
      721 gaattcgaaa acctgtattt tcagggcggg aaatcagctc tggacctggc agatccttca
      781 gcaaaagctg tcctcacagg tgaatacaaa aaagacgaac tcctagaagc tgctaggagt
      841 ggtaatgaag aaaaactaat ggctttactg actcctctaa atgtgaattg ccatgcaagt
      901 gatggacgaa agtctactcc gttacacctg gcagcaggct acaacagagt tcggatagtt
      961 cagctgcttc tacagcatgg tgctgatgtc cacgcgaaag acaaaggtgg acttgtacct
     1021 cttcataatg catgctcata cggacactat gaggtcacag aactgctgct aaagcatgga
     1081 gcttgtgtca acgctatgga tctgtggcag ttcacaccgc tgcatgaagc tgcttcaaag
     1141 aaccgggtag aagtctgttc tctgttactt agtcacggtg ccgatccaac cttggtcaat
     1201 tgccatggca aaagtgctgt cgacatggct ccaactcctg agctccggga aagactgaca
     1261 tatgaattta aaggccattc tttactccaa gccgccagag aagcagacct ggccaaagtt
     1321 aaaaaaacgc ttgctttgga aatcattaac ttcaagcagc cacagtctca tgaaactgca
     1381 ttgcactgtg ctgtggcctc cttgcatccc aaacgaaagc aggtggcaga gctgttactg
     1441 aggaagggag ccaacgtcaa cgaaaagaac aaggatttca tgacccctct gcatgttgca
     1501 gctgaaagag ctcataatga tgtcatggaa gttctgcata agcacggagc aaagatgaat
     1561 gcactggata gtcttggtca gactgctttg cacagagctg ccttagcggg ccacctgcag
     1621 acctgccgcc tcctgctgag ttatggctct gatccttcca tcatctccct gcaaggcttc
     1681 acggcagcgc agatggggaa tgaagcagtg caacagattc tgagcgagag tactcctatg
     1741 cgcacttcgg acgtggacta tcgtctctta gaggcatcga aagctggtga ctaggaaact
     1801 gtgaagcaac tctgcagccc gcagaatgtg aactgcagag acctggaggg ccggcactct
     1861 acacccttac acttcgctgc aggctataac cgtgtgtctg tcgttgagta ccttctacat
     1921 catggagctg atgtccatgc caaagacaaa ggtggcttgg ttccccttca caatgcctgc
     1981 tcctatggac actatgaggt cgccgagctc ttggtgaggc acggggcttc agtcaatgtg
     2041 gccgacctgt ggaaattcac ccctctccat gaagcggcag ccaagggaaa atacgaaatc
     2101 tgcaaactcc ttctaaaaca tggagcagat ccaactaaga agaacagaga tggaaataca
     2161 cccctagatc tggtcaaaga aggagacacg gatattcagg acctgctgag gggcgattaa
     2221 gccctgttgg atgctgccaa gaagggctgc ctagccagag tgcagaagct ttgtactcca
     2281 gagaatatca actgccgaga cacccagggc agaaattcaa cccctctcca cctggcagca
     2341 ggctacaata acctggaagt agctgagtac cttctagagc acggggctga tgtcaatgcc
     2401 caggacaaag gagggctaat cccacttcac aacgctgcgt cttatgggca tgttgacata
     2461 gcagcgttac tgataaaata caacacgtgt gtaaacgcaa cggataagtg ggcctttact
     2521 ccccttcacg aagctgccca gaaaggaagg acgcagttat gtgccctgct cctagcacat
     2581 ggcgcagacc caaccatgaa gaaccaggaa ggtcagacac ctttagatct ggcgacggct
     2641 gatgacatcc gggctctgct gatagatgcc atgccccctg aggccttacc tacctgtttt
     2701 aaacctcagt gagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
     2761 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2821 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2881 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
     2941 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3001 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3361 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3661 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4921 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4981 ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     5041 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     5101 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     5161 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     5221 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     5281 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     5341 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     5401 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     5461 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     5521 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     5581 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     5641 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     5701 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     5761 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     5821 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     5881 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     5941 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     6001 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     6061 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6121 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     6181 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     6241 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg
     6301 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     6361 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     6421 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
     6481 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
     6541 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     6601 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     6661 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga
     6721 attagcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct
     6781 gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc
     6841 cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg
     6901 agctgttgac aattaatcat cggctcgtat aatgtgtgga a
//
