LOCUS       sequence-142611-        5502 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 5502)
  TITLE     human peroxisome proliferator activated receptor alpha A/B d
REFERENCE   2  (bases 1 to 5502)
  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"
COMMENT     Sequence Label: pTrcHisA-PPARalpha(aa101-468)
FEATURES             Location/Qualifiers
     source          1..5502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             446..478
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(482..490,491..505)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1685..1702)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1855..1941
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2033..2060
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2080..2171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2172..2240,2241..3032)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2390..2409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3203..3791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3692..3711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3945..3962
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3977..4117
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4043..4062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4203..4225
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4303..4380
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4381..5463
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4400..4419)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gacgatgacg ataaggatcg atggggatcc gaatgccgta tctgcggtga
      541 caaagcttct ggttaccact acggtgttca cgcttgcgaa ggttgcaaag gtttcttccg
      601 tcgtaccatc cgtctgaaac tggtttacga caaatgcgac cgttcttgca aaatccagaa
      661 aaaaaaccgt aacaaatgcc agtactgccg tttccacaaa tgcctgtctg ttggtatgtc
      721 tcacaacgct atccgtttcg gtcgtatgcc gcgttctgaa aaagctaaac tgaaagctga
      781 aatcctgacc tgcgaacacg acatcgaaga ctctgaaacc gctgacctga aatctctggc
      841 taaacgtatc tacgaagctt acctgaaaaa cttcaacatg aacaaagtta aagctcgtgt
      901 tatcctgtct ggtaaagctt ctaacaaccc gccgttcgtt atccacgaca tggaaaccct
      961 gtgcatggct gaaaaaaccc tggttgctaa actggttgct aacggtatcc agaacaaaga
     1021 agctgaagtt cgtatcttcc actgctgcca gtgcacctct gttgaaaccg ttaccgaact
     1081 gaccgaattc gctaaagcta tcccgggttt cgctaacctg gacctgaacg accaggttac
     1141 cctgctgaaa tacggtgttt acgaagctat cttcgctatg ctgtcttctg ttatgaacaa
     1201 agacggtatg ctggttgctt acggtaacgg tttcatcacc cgtgaattcc tgaaatctct
     1261 gcgtaaaccg ttctgcgaca tcatggaacc gaaattcgac ttcgctatga aattcaacgc
     1321 tctggaactg gacgactctg acatctctct gttcgttgct gctatcatct gctgcggtga
     1381 ccgtccgggt ctgctgaacg ttggtcacat cgaaaaaatg caggaaggta tcgttcacgt
     1441 tctgcgtctg cacctgcagt ctaaccaccc ggacgacatc ttcctgttcc cgaaactgct
     1501 gcagaaaatg gctgacctgc gtcagctggt taccgaacac gctcagctgg ttcagatcat
     1561 caaaaaaacc gaatctgacg ctgctctgca cccgctgctg caggaaatct accgtgacat
     1621 gtactagggt accatatggg aattcgaagc ttggctgttt tggcggatga gagaagattt
     1681 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     1741 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     1801 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     1861 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1921 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     1981 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     2041 atcctgacgg atggcctttt tgcgtttcta caaactcttt ttgtttattt ttctaaatac
     2101 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2161 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2221 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2281 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2341 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2401 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2461 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2521 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2581 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2641 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2701 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2761 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2821 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2881 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2941 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3001 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3061 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3121 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3781 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3841 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3901 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3961 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4021 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4081 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4141 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4201 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca
     4261 gatcaattcg cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggcgc
     4321 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     4381 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     4441 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     4501 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     4561 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     4621 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     4681 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     4741 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     4801 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     4861 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     4921 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     4981 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     5041 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     5101 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     5161 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     5221 tgttatatcc cgccgtcaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     5281 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     5341 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     5401 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5461 tgagcgcaac gcaattaatg tgagttagcg cgaattgatc tg
//
