LOCUS       sequence-415029-        5718 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 5718)
  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 5718)
  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-PPARdelta
FEATURES             Location/Qualifiers
     source          1..5718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(105..110,111..126,127..133)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    141..157
                     /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             180..833
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     791..813
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             840..857
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1690..1712)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1857..1875)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1943..2047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2048..2116,2117..2908)
                     /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(2266..2285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3079..3667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3568..3587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3821..3838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3911..3988
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3989..5071
                     /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(4008..4027)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5120..5125,5126..5143,5144..5150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5158..5174
                     /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     5182..5198
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5182..5198
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5194..5718,1..3)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5214..5231)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5214..5230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5223..5245)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
       61 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
      121 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca
      181 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
      241 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
      301 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
      361 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
      421 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
      481 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
      541 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
      601 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
      661 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
      721 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
      781 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
      841 tggttccgcg tggatctcgt cgtgcatctg ttggatccat ggagcagcca caggaggaag
      901 ctcctgaggt ccgggaagag gaggagaaag aggaagtggc agaggcagaa ggagccccag
      961 agctcaatgg gggaccacag catgcacttc cttccagcag ctacacagac ctctcccgga
     1021 gctcctcgcc accctcactg ctggaccaac tgcagatggg ctgtgacggg gcctcatgcg
     1081 gcagcctcaa catggagtgc cgggtgtgcg gggacaaggc atcgggcttc cactacggtg
     1141 ttcatgcatg tgaggggtgc aagggcttct tccgtcgtac gatccgcatg aagctggagt
     1201 acgagaagtg tgagcgcagc tgcaagattc agaagaagaa ccgcaacaag tgccagtact
     1261 gccgcttcca gaagtgcctg gcactgggca tgtcacacaa cgctatccgt tttggtcgga
     1321 tgccggaggc tgagaagagg aagctggtgg cggggctgac tgcaaatgag gggagccagt
     1381 acaacccaca ggtggccgac ctgaaggcct tctccaagca catctacaat gcctacctga
     1441 aaaacttcag catgaccaaa aagaaggccc gcagcatcct caccggcaaa gccagccaca
     1501 cggcgccctt tgtgatccac gacatcgaga cattgtggca ggcagagaag gggctggtgt
     1561 ggaagcagtt ggtgaatggc ctgcctccct acaaggagat cagcgtgcac gccttctacc
     1621 gctgccagta agaattcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga
     1681 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     1741 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     1801 agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac gaaagggcct
     1861 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1921 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1981 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2041 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2101 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2161 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2221 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2281 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2341 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2401 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2461 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2521 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2581 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2641 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2701 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2761 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2821 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2881 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2941 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3001 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3061 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3121 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3181 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3241 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3301 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3361 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3421 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3481 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3541 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3601 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3661 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3721 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3781 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3841 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3901 cataaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     3961 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     4021 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     4081 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     4141 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     4201 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     4261 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     4321 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     4381 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     4441 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     4501 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     4561 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     4621 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     4681 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     4741 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     4801 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     4861 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     4921 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     4981 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5041 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     5101 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5161 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc
     5221 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     5281 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     5341 caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg
     5401 gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca
     5461 aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg
     5521 gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat
     5581 gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt
     5641 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
     5701 gtatggctgt gcaggtcg
//
