LOCUS       sequence-415027-        5727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5727)
  AUTHORS   He TC, Chan TA, Vogelstein B, Kinzler KW
  TITLE     PPARdelta is an APC-regulated target of nonsteroidal 
            anti-inflammatory drugs.
  JOURNAL   Cell. 1999 Oct 29. 99(3):335-45.
  PUBMED    10555149
REFERENCE   2  (bases 1 to 5727)
  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: "Cell. 1999 
            Oct 29. 99(3):335-45."
COMMENT     Sequence Label: pGST-PPARgamma
FEATURES             Location/Qualifiers
     source          1..5727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(56..61,62..77,78..84)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    92..108
                     /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             131..784
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     742..764
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             791..808
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1650..1672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1817..1835)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1903..2007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2008..2076,2077..2868)
                     /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(2226..2245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3039..3627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3528..3547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3781..3798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3871..3948
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3949..5031
                     /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(3968..3987)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5080..5085,5086..5103,5104..5110)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5118..5134
                     /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     5142..5158
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5142..5158
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5154..5681
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5174..5191)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5174..5190)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5183..5205)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
       61 aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
      121 aacagtattc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
      181 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
      241 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
      301 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
      361 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
      421 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
      481 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
      541 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
      601 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
      661 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
      721 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
      781 aaaatcggat ctggttccgc gtggatctcg tcgtgcatct gttggatccc cgggaattcc
      841 gatgaccatg gttgacacag tgattccatt ctggcccacc aactttggga tcagctccgt
      901 ggatctctcc gtaatggaag accactccca ctcctttgat atcaagccct tcactactgt
      961 tgacttctcc agcatttcta ctccacatta cgaagacatt ccatacacaa gaacagatcc
     1021 agtggttgca gattacaagt atgacctgaa acttcaagag taccaaagtg caatcaaagt
     1081 ggagcctgca tctccacctt attattctga gaagactcag ctctacaata agcctcatga
     1141 agagccttcc aactccctca tggcaattga atgtcgtgtc tgtggagata gagcttctgg
     1201 atttcactat ggagttcatg cttgtgaagg atgcaagggt ttcttccgga gaacaatcag
     1261 attgaagctt atctatgaca gatgtgatct taactgtcgg atccacaaaa aaagtagaaa
     1321 taaatgtcag tactgtcggt ttcagaaatg ccttgcagtg gggatgtctc ataatgccat
     1381 caggtttggg cggatgccac aggccgagaa ggagaagctg ttggcggaga tctccagtga
     1441 tatcgaccag ctgaatccag agtccgctga cctccaggcc ctggcaaaac atttgtatga
     1501 ctcgtacata aagtccttcc cgctgaccaa agcaaaggcg agggcgatct tgacaggaaa
     1561 gacaacagac aaatcaccat tcgttatcta agaattcatc gtgactgact gacgatctgc
     1621 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1681 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1741 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat
     1801 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     1861 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1921 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1981 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2041 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2101 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2161 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2221 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2281 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2341 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2401 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2461 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2521 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     2581 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2641 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2701 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2761 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2821 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2881 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2941 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3001 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3061 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3181 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3721 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3781 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3841 ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggcgcaa aacctttcgc
     3901 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     3961 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     4021 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     4081 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     4141 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     4201 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     4261 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     4321 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     4381 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     4441 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     4501 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     4561 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     4621 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     4681 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     4741 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     4801 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     4861 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     4921 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     4981 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5041 aattaatgta agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5101 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5161 tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     5221 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     5281 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct
     5341 ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg
     5401 atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca
     5461 acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt
     5521 gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta
     5581 tttttgatgg cgttggaatt agcttatcga ctgcacggtg caccaatgct tctggcgtca
     5641 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     5701 cgctcaaggc gcactcccgt tctggat
//
