LOCUS       sequence-469916-        6384 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Protein overexpression of human uroporphyrinogen decarboxylase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6384)
  TITLE     A method for the detection and enrichment of endogenous cereblon 
            substrates
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6384)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET28a-His6-UROD
FEATURES             Location/Qualifiers
     source          1..6384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..91
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(111..188)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     394..413
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     497..516
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        497..515
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    516..540
                     /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)."
     RBS             555..577
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             588..605
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             630..650
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1815..1833)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1829..1876
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1913..2368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2000..2019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2210..2231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2460..3275)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2460..3275)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3183..3202
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3267..3286
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3397..3985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3886..3905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4139..4156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4171..4310
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4236..4255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4396..4418
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4412..4603)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5412..6384,1..110))
                     /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"
ORIGIN
        1 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
       61 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
      121 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
      181 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
      241 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
      301 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
      361 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
      421 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
      481 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
      541 cctctagaaa taattttgtt taactttaag aaggagatat acatatgcat caccatcacc
      601 accattcgtc cggagtcgac ttaggcaccg agaatttgta ttttcagagc atggaagcga
      661 acggccttgg ccctcaaggg tttccagagt tgaaaaacga taccttcctt cgtgcagcct
      721 ggggggaaga aacggactat acacccgtgt ggtgtatgcg ccaggctggc cgttaccttc
      781 cagagttccg cgaaacgcgt gcggcgcaag atttttttag cacctgccgt tctcccgagg
      841 cgtgctgcga attgactctt cagccgttac gccgttttcc acttgatgcg gcgatcatct
      901 tcagcgacat tttggtagtc cctcaagctc tgggtatgga ggtaacgatg gttccaggga
      961 aggggccgtc ctttccagaa ccactgcgcg aagagcagga cttggaacgc ttgcgtgacc
     1021 cggaggtggt ggcgtccgag ctggggtatg tcttccaggc tattaccctt acacgccagc
     1081 gtcttgcggg acgtgttcct ctgattggat ttgctggagc accatggact cttatgacct
     1141 acatggtaga gggaggaggc agctctacta tggcccaggc aaaacgctgg ttgtaccaac
     1201 gtccacaagc gtcgcaccaa ttgttacgca ttctgaccga tgccttggtg ccttatcttg
     1261 tgggtcaggt cgtcgcaggt gcgcaggcct tgcagttatt tgagtcacac gcaggtcacc
     1321 ttggaccaca gctgtttaac aaatttgctc tgccctatat ccgtgatgtg gccaagcaag
     1381 ttaaggcccg tttgcgcgag gcgggtcttg caccagtacc tatgatcatc tttgccaagg
     1441 atgggcactt cgccttggag gaattggcac aggcaggcta cgaagtagtt gggcttgact
     1501 ggacggtagc cccgaagaaa gctcgtgagt gcgtggggaa gacggttaca ttgcagggga
     1561 atctggaccc ctgtgcttta tatgctagcg aagaggaaat cggtcaactt gtcaaacaaa
     1621 tgttggacga ctttggtcca catcgctata tcgcaaatct tggccatgga ctgtacccgg
     1681 atatggatcc agagcacgta ggcgcattcg ttgatgccgt ccataagcac tcccgtttac
     1741 tgcgccaaaa ttaaaggtgg atacggatcc ggctgctaac aaagcccgaa aggaagctga
     1801 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1861 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
     1921 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1981 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2041 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2101 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2161 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2221 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2281 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2341 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     2401 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
     2461 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     2521 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     2581 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     2641 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     2701 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     2761 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     2821 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     2881 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     2941 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     3001 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     3061 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     3121 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     3181 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     3241 ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     3301 taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3541 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4021 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4081 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4141 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     4201 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4261 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     4321 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4381 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4441 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     4501 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4561 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     4621 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     4681 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     4741 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     4801 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     4861 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     4921 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     4981 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     5041 acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct
     5101 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     5161 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     5221 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     5281 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     5341 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     5401 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5461 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     5521 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     5581 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     5641 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     5701 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     5761 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     5821 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     5881 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     5941 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     6001 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     6061 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     6121 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     6181 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     6241 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     6301 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     6361 agctccgcca tcgccgcttc cact
//
