LOCUS       sequence-259514-        7352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mouse Tankyrase1 5ARC (171–961) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7352)
  AUTHORS   DaRosa PA, Klevit RE, Xu W
  TITLE     Structural basis for tankyrase-RNF146 interaction reveals 
            noncanonical tankyrase-binding motifs.
  JOURNAL   Protein Sci. 2018 Jun;27(6):1057-1067. doi: 10.1002/pro.3413. Epub 
            2018 Apr 25.
  PUBMED    29604130
REFERENCE   2  (bases 1 to 7352)
  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: "10.1002/pro.3413"; 
            journalName: "Protein Sci"; date: "2018-06"; volume: "27"; issue: 
            "6"; pages: "1057-1067"
COMMENT     Sequence Label: pGEX-4T1 mouse Tankyrase1 5ARC (171–961)
FEATURES             Location/Qualifiers
     source          1..7352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    34..50
                     /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             73..726
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     684..706
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             733..750
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             760..780
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3217..3239)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3384..3402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3470..3574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3575..3643,3644..4435)
                     /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(3793..3812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4606..5194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5095..5114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5348..5365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5438..5515
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5516..6598
                     /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(5535..5554)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6647..6652,6653..6670,6671..6677)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6685..6701
                     /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     6709..6725
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6709..6725
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6721..7248
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6741..6758)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6741..6757)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6750..6772)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(7350..7352,1..3,4..19,20..26)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
       61 gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc
      121 actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat
      181 gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct
      241 tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct
      301 gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa
      361 ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa
      421 actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt
      481 ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat
      541 gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta
      601 gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc
      661 aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct
      721 ccaaaatcgg atctggttcc gcgtggatcc ccggaattcg aaaacctgta ttttcaaggt
      781 gccgtgagtg gggccctgcg ggagctgctc gaggcctgtc ggaatgggga cgtgtcccgg
      841 gtgaagaggt tggtggacgc cgcgaacgtg aatgccaagg acatggccgg ccgaaagtct
      901 tctcccctgc acttcgcagc aggttttgga aggaaggatg ttgtagaaca cttgctgcag
      961 atgggtgcta atgtccacgc ccgtgatgac gggggtctca tcccacttca taatgcctgc
     1021 tctttcggcc atgctgaggt tgtgagtctg ttactgtgtc aaggagcaga tccgaatgcc
     1081 agagataact ggaactatac accactgcat gaagctgcca ttaaagggaa aattgatgtc
     1141 tgtattgtgc tgctgcagca cggagccgat ccaaacatcc ggaacaccga tgggaaatca
     1201 gctctggacc tggcagatcc ttcagcaaaa gctgtcctca caggtgaata caaaaaagac
     1261 gaactcctag aagctgctag gagtggtaat gaagaaaaac taatggcttt actgactcct
     1321 ctaaatgtga attgccatgc aagtgatgga cgaaagtcta ctccgttaca cctggcagca
     1381 ggctacaaca gagttcggat agttcagctg cttctacagc atggtgctga tgtccacgcg
     1441 aaagacaaag gtggacttgt acctcttcat aatgcatgct catacggaca ctatgaggtc
     1501 acagaactgc tgctaaagca tggagcttgt gtcaacgcta tggatctgtg gcagttcaca
     1561 ccgctgcatg aagctgcttc aaagaaccgg gtagaagtct gttctctgtt acttagtcac
     1621 ggtgccgatc caaccttggt caattgccat ggcaaaagtg ctgtcgacat ggctccaact
     1681 cctgagctcc gggaaagact gacatatgaa tttaaaggcc attctttact ccaagccgcc
     1741 agagaagcag acctggccaa agttaaaaaa acgcttgctt tggaaatcat taacttcaag
     1801 cagccacagt ctcatgaaac tgcattgcac tgtgctgtgg cctccttgca tcccaaacga
     1861 aagcaggtgg cagagctgtt actgaggaag ggagccaacg tcaacgaaaa gaacaaggat
     1921 ttcatgaccc ctctgcatgt tgcagctgaa agagctcata atgatgtcat ggaagttctg
     1981 cataagcacg gagcaaagat gaatgcactg gatagtcttg gtcagactgc tttgcacaga
     2041 gctgccttag cgggccacct gcagacctgc cgcctcctgc tgagttatgg ctctgatcct
     2101 tccatcatct ccctgcaagg cttcacggca gcgcagatgg ggaatgaagc agtgcaacag
     2161 attctgagcg agagtactcc tatgcgcact tcggacgtgg actatcgtct cttagaggca
     2221 tcgaaagctg gtgacttgga aactgtgaag caactctgca gcccgcagaa tgtgaactgc
     2281 agagacctgg agggccggca ctctacaccc ttacacttcg ctgcaggcta taaccgtgtg
     2341 tctgtcgttg agtaccttct acatcatgga gctgatgtcc atgccaaaga caaaggtggc
     2401 ttggttcccc ttcacaatgc ctgctcctat ggacactatg aggtcgccga gctcttggtg
     2461 aggcacgggg cttcagtcaa tgtggccgac ctgtggaaat tcacccctct ccatgaagcg
     2521 gcagccaagg gaaaatacga aatctgcaaa ctccttctaa aacatggagc agatccaact
     2581 aagaagaaca gagatggaaa tacaccccta gatctggtca aagaaggaga cacggatatt
     2641 caggacctgc tgaggggcga tgctgccctg ttggatgctg ccaagaaggg ctgcctagcc
     2701 agagtgcaga agctttgtac tccagagaat atcaactgcc gagacaccca gggcagaaat
     2761 tcaacccctc tccacctggc agcaggctac aataacctgg aagtagctga gtaccttcta
     2821 gagcacgggg ctgatgtcaa tgcccaggac aaaggagggc taatcccact tcacaacgct
     2881 gcgtcttatg ggcatgttga catagcagcg ttactgataa aatacaacac gtgtgtaaac
     2941 gcaacggata agtgggcctt tactcccctt cacgaagctg cccagaaagg aaggacgcag
     3001 ttatgtgccc tgctcctagc acatggcgca gacccaacca tgaagaacca ggaaggtcag
     3061 acacctttag atctggcgac ggctgatgac atccgggctc tgctgataga tgccatgccc
     3121 cctgaggcct tacctacctg ttttaaacct cagtgagcgg ccgcatcgtg actgactgac
     3181 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3241 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3301 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     3361 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3421 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3481 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3541 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3601 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3661 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3721 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3781 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     3841 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3901 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3961 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4021 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4081 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     4141 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4201 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4261 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4321 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4381 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4441 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4501 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4561 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4621 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4681 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4741 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4801 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4861 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4921 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4981 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5041 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5101 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5161 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5221 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5281 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5341 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     5401 tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat ggcgcaaaac
     5461 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     5521 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     5581 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     5641 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     5701 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     5761 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     5821 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     5881 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     5941 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     6001 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     6061 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     6121 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     6181 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     6241 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     6301 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     6361 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     6421 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     6481 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     6541 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     6601 gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta cactttatgc
     6661 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     6721 atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6781 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6841 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     6901 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     6961 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     7021 tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     7081 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     7141 cgaattattt ttgatggcgt tggaattagc ttatcgactg cacggtgcac caatgcttct
     7201 ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa
     7261 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     7321 ggttctggca aatattctga aatgagctgt tg
//
