LOCUS       sequence-446475-        5488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-tagged human PHD2 catalytic domain with TEV cleavage site 
            between His and PHD2 for PHD2 purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5488)
  AUTHORS   Gurol KC, Jursa T, Cho EJ, Fast W, Dalby KN, Smith DR, Mukhopadhyay 
            S
  TITLE     PHD2 enzyme is an intracellular manganese sensor that initiates the 
            homeostatic response against elevated manganese.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Jun 25;121(26):e2402538121. doi: 
            10.1073/pnas.2402538121. Epub 2024 Jun 21.
  PUBMED    38905240
REFERENCE   2  (bases 1 to 5488)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-06-25"; volume: "121(26):e2402538121. 
            doi"; pages: " 10.1073/pnas.2402538121. Epub 2024 Jun 21"
COMMENT     Sequence Label: His-PHD2-181-426
FEATURES             Location/Qualifiers
     source          1..5488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      62..89
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        108..199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(200..268,269..1060)
                     /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(418..437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1231..1819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1720..1739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1973..1990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2067..2495)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2177..2198)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2389..2408
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2533..2552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2693..2715
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2793..2870
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2871..3953
                     /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(2890..2909)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    4222..4238
                     /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     4227..4249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4271..4288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4310..4330
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5202..5219)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5372..5458
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
       61 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
      121 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      181 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      241 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      301 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      361 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      421 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
      481 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      541 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      601 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      661 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      721 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      781 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      841 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      901 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      961 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1021 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1081 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1141 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1201 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1261 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1321 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     1381 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1441 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1501 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1561 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1621 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1681 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1741 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1801 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1861 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1921 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1981 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2041 tatttcacac cgcataattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2101 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2161 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2221 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2281 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2341 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2401 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2461 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgctatg
     2521 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     2581 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     2641 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2701 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg
     2761 cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc
     2821 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     2881 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     2941 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     3001 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     3061 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     3121 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     3181 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     3241 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     3301 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     3361 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     3421 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     3481 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     3541 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     3601 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     3661 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     3721 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     3781 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     3841 tgaaaagaaa aaccaccctg gcacccaata cgcaaaccgc ctctccccgc gcgttggccg
     3901 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3961 gcaattaatg taagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg
     4021 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     4081 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     4141 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     4201 cggtccgtat aatctgtgga attgtgagcg gataacaatt tcacacagga aacagaccat
     4261 gtcgtactac catcaccatc accatcacga ttacgatatc ccaacgaccg aaaacctgta
     4321 ttttcagggc gccatggatc ccaacgggca gacgaagccc ctgccggcgc tgaagctggc
     4381 gctcgagtac atcgtgccgt gcatgaacaa gcacggcatc tgtgtggtgg acgacttcct
     4441 cggcaaggag accggacagc agatcggcga cgaggtgcgc gccctgcacg acaccgggaa
     4501 gttcacggac gggcagctgg tcagccagaa gagtgactcg tccaaggaca tccgaggcga
     4561 taagatcacc tggatcgagg gcaaggagcc cggctgcgaa accattgggc tgctcatgag
     4621 cagcatggac gacctgatac gccactgtaa cgggaagctg ggcagctaca aaatcaatgg
     4681 ccggacgaaa gccatggttg cttgttatcc gggcaatgga acgggttatg tacgtcatgt
     4741 tgataatcca aatggagatg gaagatgtgt gacatgtata tattatctta ataaagactg
     4801 ggatgccaag gtaagtggag gtatacttcg aatttttcca gaaggcaaag cccagtttgc
     4861 tgacattgaa cccaaatttg atagactgct gtttttctgg tctgaccgtc gcaaccctca
     4921 tgaagtacaa ccagcatatg ctacaaggta cgcaataact gtttggtatt ttgatgcaga
     4981 tgagagagca cgagctaaag taaaatatct aacaggtgaa aaaggtgtga gggttgaact
     5041 caataaacct tcagattcgg tcggtaaaga cgtcttctag gaattcaaag gcctacgtcg
     5101 acgagctcaa ctagtgcggc cgctttcgaa tctagagcct gcagtctcga ggcatgcggt
     5161 accaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     5221 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     5281 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     5341 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     5401 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     5461 gccgggagcg gatttgaacg ttgcgaag
//
