LOCUS       sequence-389117-        6643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses tagless mouse ZNG1 in the pHIS vector (inserted at 
            NdeI-BamHI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6643)
  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 6643)
  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: mouse ZNG1
FEATURES             Location/Qualifiers
     source          1..6643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      24..479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(111..130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     321..342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        505..609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(610..678,679..1470)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(828..847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1641..2229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2130..2149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2383..2400
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2415..2554
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2480..2499)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2640..2662
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2656..2847)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3656..4738)
                     /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     4700..4719
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4739..4816)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5022..5041
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5125..5144
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5125..5143
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5144..5168
                     /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             5183..5205
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6499..6516
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6569..6587)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6583..6630
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
       61 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
      121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      181 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
      241 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      301 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
      361 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
      421 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
      481 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      541 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      601 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      661 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      721 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      781 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      841 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      901 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
      961 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1021 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1081 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1141 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1201 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1261 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1321 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1381 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1441 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1501 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1561 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1621 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1681 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1741 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     1801 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1861 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1921 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1981 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2041 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2101 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2161 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2221 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2281 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2341 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2401 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2461 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     2521 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     2581 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2641 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     2701 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     2761 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     2821 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     2881 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     2941 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3001 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     3061 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     3121 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     3181 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     3241 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     3301 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     3361 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     3421 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     3481 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     3541 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     3601 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     3661 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3721 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     3781 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     3841 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     3901 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     3961 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     4021 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     4081 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     4141 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     4201 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     4261 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     4321 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     4381 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     4441 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     4501 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     4561 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     4621 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     4681 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     4741 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     4801 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     4861 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     4921 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     4981 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     5041 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     5101 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     5161 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac atatgttacc
     5221 tgctgtgaaa actgtggagg cggaagaaga gtacgcggag gattgtcctg aattggtccc
     5281 cattgagacc aagaaccaag aagaggaaaa tcttgacttc atcactaaga ttccagtcac
     5341 aattgtcacc gggtacttag gtgctggaaa gacaacactt ctgaactaca ttttgacaga
     5401 acaacataat agaaaaatag cagtgatttt aaatgaattt ggagaaggga gtgcagtaga
     5461 gaaatcctta gctgtcagtc aaggtggaga actctatgaa gaatggctgg aacttagaaa
     5521 tggttgcctc tgctgttcag tcaaggacag tggccttaga gctatagaga atctgatgca
     5581 aaagaagggc aagtttgact acatactctt agagaccact gggctagcag atcctggtgc
     5641 agtggcttct atgttttggg ttgatgctga gttagggagt gatatttatc ttgatggtat
     5701 cataactgtt gtagattcaa aatatggatt aaaacattta actgaagaaa aacctgatgg
     5761 ccttgtcaat gaagctacta ggcaagttgc attggcagac atgattctca tcaataaaac
     5821 agacttggtt tcagaagagg agctaaacaa cttaagaaca actatcagat caataaatgg
     5881 actgggaaaa gttttagaaa ctcagagatc aagagttcat ctctcaaata tattagacct
     5941 tcatgcctat gatattctct ctggaataag tttacagaaa aagctccagc atgtgtcaac
     6001 agcaccccac cttgatcaga gtattgttac agtcacattt gaagtaccag gaagtgcaaa
     6061 ggaagaatgt ctaaatgtat ttatccagaa tctcctgtgg gaaaagaacg tgaaaaacaa
     6121 ggacggccac tgcatggagg tcattcggct aaagggactg gtgtctatca aagacaaacc
     6181 acagcaaatg attgtgcaag gcatccatga gctgtacgac ctggaagaaa gcctagtgaa
     6241 ctggaaggat gatgcggaga gagcgtgtca gctggtcttc attggtagaa atttagataa
     6301 ggatgttctt caacaactct tcctcactgc tgtggcagag gcagaagagc aaaggacagc
     6361 accaggcaga gatggagttt gtccatcgca ctaggaattc aaaggcctac gtcgacgagc
     6421 tcaactagtg cggccgcttt cgaatctaga gcctgcagtc tcgaggcatg cggtaccaag
     6481 cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     6541 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     6601 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaa
//
