LOCUS       sequence-389113-        5805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a SUMO-tagged fusion of ZNG1, residues 10-30 and mouse 
            METAP1, residues 1-59.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5805)
  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 5805)
  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 ZNG1-METAP1 fusion
FEATURES             Location/Qualifiers
     source          1..5805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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             79..101
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             114..131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             135..425
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             690..707
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(760..778)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      774..821
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      858..1313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(945..964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1155..1176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1405..2220)
                     /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(1405..2220)
                     /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     2128..2147
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2212..2231
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2342..2930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2831..2850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3084..3101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3116..3255
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3181..3200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3341..3363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3357..3548)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4357..5439)
                     /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     5401..5420
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5440..5517)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5723..5742
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggtcatcacc
      121 atcatcatca cgggtcggac tcagaagtca atcaagaagc taagccagag gtcaagccag
      181 aagtcaagcc tgagactcac atcaatttaa aggtgtccga tggatcttca gagatcttct
      241 tcaagatcaa aaagaccact cctttaagaa ggctgatgga agcgttcgct aaaagacagg
      301 gtaaggaaat ggactcctta agattcttgt acgacggtat tagaattcaa gctgatcagg
      361 cccctgaaga tttggacatg gaggataacg atattattga ggctcaccgc gaacagattg
      421 gaggagcgga agaagagtac gcggaggatt gtcctgaatt ggtccccatt gagaccaaga
      481 accaagaaat ggcggcggtg gagacgcggg tgtgcgagac ggacggctgc agcagcgagg
      541 ccaagctcca gtgtcccacc tgcatcaagc tgggcatcca gggctcgtac ttctgctcgc
      601 aggaatgctt taaaggaagc tgggccactc acaagttact acataagaaa gcaaaagatg
      661 aaaagtaaaa gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg
      721 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
      781 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
      841 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      901 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      961 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1021 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1081 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1141 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1201 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     1261 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     1321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1381 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     1441 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     1501 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     1561 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     1621 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     1681 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     1741 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     1801 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     1861 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     1921 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     1981 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     2041 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     2101 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     2161 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     2221 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc
     2281 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2341 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2401 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2461 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2521 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2581 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2641 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2701 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2761 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2821 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2881 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2941 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3001 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3061 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3121 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3181 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3241 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3301 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3361 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3421 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3481 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     3541 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     3601 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     3661 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     3721 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     3781 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     3841 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     3901 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     3961 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4021 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4081 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4141 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4201 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4261 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     4321 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4381 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4441 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     4501 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     4561 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     4621 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     4681 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     4741 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     4801 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     4861 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     4921 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     4981 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5041 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5101 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5161 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5221 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     5281 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     5341 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     5401 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     5461 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     5521 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     5581 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     5641 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     5701 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     5761 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcg
//
