LOCUS       sequence-389112-        5748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SUMO-tagged mouse METAP1, residues 1-59.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5748)
  AUTHORS   Weiss A, Murdoch CC, Edmonds KA, Jordan MR, Monteith AJ, Perera YR, 
            Rodriguez Nassif AM, Petoletti AM, Beavers WN, Munneke MJ, Drury SL,
            Krystofiak ES, Thalluri K, Wu H, Kruse ARS, DiMarchi RD, Caprioli 
            RM, Spraggins JM, Chazin WJ, Giedroc DP, Skaar EP
  TITLE     Zn-regulated GTPase metalloprotein activator 1 modulates vertebrate 
            zinc homeostasis.
  JOURNAL   Cell. 2022 Jun 9;185(12):2148-2163.e27. doi: 
            10.1016/j.cell.2022.04.011. Epub 2022 May 17.
  PUBMED    35584702
REFERENCE   2  (bases 1 to 5748)
  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"; doi: 
            "10.1016/j.cell.2022.04.011"; journalName: "Cell"; date: "2022-06-9-
            9"; volume: "185"; issue: "12"; pages: "2148-2163.e27"
COMMENT     Sequence Label: pSUMO mouse METAP1 1-59
FEATURES             Location/Qualifiers
     source          1..5748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    42..66
                     /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             81..103
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             116..133
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             137..427
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             635..652
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(705..723)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      719..766
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      803..1258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(890..909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1100..1121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1350..2165)
                     /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(1350..2165)
                     /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     2073..2092
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2157..2176
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2287..2875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2776..2795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3029..3046
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3061..3200
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3126..3145)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3286..3308
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3302..3493)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4302..5384)
                     /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     5346..5365
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5385..5462)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5668..5687
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
       61 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tgggtcatca
      121 ccatcatcat cacgggtcgg actcagaagt caatcaagaa gctaagccag aggtcaagcc
      181 agaagtcaag cctgagactc acatcaattt aaaggtgtcc gatggatctt cagagatctt
      241 cttcaagatc aaaaagacca ctcctttaag aaggctgatg gaagcgttcg ctaaaagaca
      301 gggtaaggaa atggactcct taagattctt gtacgacggt attagaattc aagctgatca
      361 ggcccctgaa gatttggaca tggaggataa cgatattatt gaggctcacc gcgaacagat
      421 tggaggatcc catatggcgg cggtggagac gcgggtgtgc gagacggacg gctgcagcag
      481 cgaggccaag ctccagtgtc ccacctgcat caagctgggc atccagggct cgtacttctg
      541 ctcgcaggaa tgctttaaag gaagctgggc cactcacaag ttactacata agaaagcaaa
      601 agatgaaaag taaaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
      661 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
      721 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
      781 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      841 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      901 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      961 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1021 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1081 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1141 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1201 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1321 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     1381 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1441 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1501 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1561 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     1621 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1681 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1741 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1801 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1861 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1921 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1981 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     2041 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     2101 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     2161 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     2221 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2281 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2341 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2401 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2461 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2521 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2581 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2641 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2701 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2761 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2821 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2881 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2941 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3001 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3061 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3121 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3181 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3241 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3301 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3361 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3421 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3481 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3541 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3601 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3661 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     3721 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     3781 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     3841 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     3901 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     3961 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     4021 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     4081 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     4141 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     4201 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     4261 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4321 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4381 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     4441 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     4501 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     4561 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     4621 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     4681 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     4741 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     4801 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     4861 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     4921 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     4981 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5041 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5101 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5161 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5221 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5281 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5341 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     5401 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     5461 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     5521 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     5581 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     5641 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     5701 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggat
//
