LOCUS       sequence-158894-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged SLC30A10-D248A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  AUTHORS   Zogzas CE, Aschner M, Mukhopadhyay S
  TITLE     Structural Elements in the Transmembrane and Cytoplasmic Domains of 
            the Metal Transporter SLC30A10 Are Required for Its Manganese Efflux
            Activity.
  JOURNAL   J Biol Chem. 2016 Jul 29;291(31):15940-57. doi: 
            10.1074/jbc.M116.726935. Epub 2016 Jun 15.
  PUBMED    27307044
REFERENCE   2  (bases 1 to 7795)
  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: 
            "10.1074/jbc.M116.726935"; journalName: "J Biol Chem"; date: 
            "2016-07-29- 29"; volume: "291"; issue: "31"; pages: "15940-57"
COMMENT     Sequence Label: FLAG-SLC30A10-D248A
FEATURES             Location/Qualifiers
     source          1..7795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2554..3176
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2753..2772)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3200..3220)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3218..3515
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3569..4363
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3623..3642)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4233..4252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5015..5149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5052..5071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5106..5125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5154..5173)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5155..5173)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5187..5203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5187..5203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5200..5222)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5211..5227
                     /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)."
     promoter        complement(join(5235..5241,5242..5259,5260..5265))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5280..5301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5418..5435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5589..6177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5669..5688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6348..7139,7140..7208))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6971..6990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7209..7313)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7340..7768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7427..7446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7637..7658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcgatgggc
     1021 cgctactctg gcaagacgtg ccggctgctc ttcatgctgg tgctcaccgt cgccttcttc
     1081 gtggcggagc tggtctccgg ctacctgggc aactccatcg cgctgctctc cgactccttc
     1141 aacatgctct ccgacctgat ctcgctgtgc gtgggcctga gcgccggcta catcgcccgg
     1201 cgccccaccc ggggcttcag cgccacctac ggctacgccc gcgccgaggt ggtgggcgcg
     1261 ctgagcaacg cggtcttcct caccgcgctc tgcttcacca tcttcgtgga ggccgtgctg
     1321 cgcctggccc ggcccgagcg catcgatgac cccgagctgg tgctcatcgt cggcgtcctg
     1381 gggctgttgg tcaacgtggt ggggctgctc atcttccagg actgcgccgc ctggttcgcg
     1441 tgctgcctcc ggggacgcag tcgccgcctg cagcagcggc agcagctggc ggagggctgt
     1501 gtccccggcg ctttcggggg gcctcagggc gcggaggacc cgcggcgcgc ggcggacccg
     1561 acagccccag gctcggactc ggccgtaacc ctccggggga cctcggtgga aaggaagcgg
     1621 gagaaggggg cgaccgtgtt cgcaaacgta gcaggtgatt ccttcaacac ccagaatgag
     1681 ccagaagaca tgatgaaaaa agagaaaaag tctgaagctc tgaatatcag aggagtactt
     1741 ttgcatgtga tgggagctgc cctggggtcc gtggttgtgg tcatcacggc catcatattc
     1801 tatgtgcttc ccctgaagag tgaggacccg tgtaactggc agtgttacat tgaccccagc
     1861 ctgactgtcc tcatggtcat catcattttg tcatctgcct tcccgcttat caaggagacc
     1921 gctgccattc tgctacagat ggtcccaaaa ggagtcaaca tggaagagct gatgagtaaa
     1981 ctctctgctg tgcctggaat tagcagtgta catgaagtgc acatctggga acttgtaagt
     2041 ggaaagatta ttgccaccct gcacatcaag tatcctaagg acaggggata tcaagatgcc
     2101 agcacaaaaa ttcgagaaat cttccaccat gcgggaatcc acaatgtgac catccagttt
     2161 gaaaatgtgg acttgaagga acccctggag cagaaggact tactgttgct ctgcaactca
     2221 ccctgcatct ccaagggctg tgctaagcag ctgtgttgtc cccccggggc actgcctctg
     2281 gctcacgtca atggctgtgc tgagcacaat ggtgggccct ctctagacac atacggaagt
     2341 gatggcctca gtagaagaga cgcaagagaa gtggctattg aagtgtcttt ggatagctgt
     2401 ctgagtgacc acggacaaag tcttaacaaa actcaggagg accaatgtta tgtcaacaga
     2461 acgcattttc ctaggtttaa gggcgaattc cagcacactg gcggccgtta ctagtggatc
     2521 tgatatcggt accagtcgac tctagaggat cccgggtggc atccctgtga cccctcccca
     2581 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     2641 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     2701 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     2761 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     2821 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     2881 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     2941 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     3001 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     3061 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     3121 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     3181 acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3481 gagctattcc agaagtagtg aggaggcttt tttggaggaa ttgatcagct tgggatctga
     3541 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3601 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3661 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3721 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3781 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3841 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3901 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3961 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4021 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4081 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4141 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4201 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4261 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4321 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4381 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4441 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4501 cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct cgcgagttgg
     4561 ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg caaatccgtc
     4621 ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact gcaggagtgg
     4681 ggaggcacga tggccgcttt ggtcgacccg gacgggacgc tcctgcgcct gatacagaac
     4741 gaattgcttg caggcatctc atgagtgtgt cttcccgttt tccgcctgag gtcactgcgt
     4801 ggatggagcg ctggcgcctg ctgcgcgacg gcgagctgct caccacccac tcgccaagct
     4861 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgccga tcataatcag
     4921 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4981 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5041 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5101 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca attccctata
     5161 gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     5221 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5281 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5341 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5401 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5461 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5521 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5581 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5641 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5701 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5761 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5821 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5881 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5941 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6001 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6061 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6121 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6181 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6241 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6361 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6421 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6481 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6541 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6601 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6661 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6721 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6781 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6841 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6901 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6961 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7021 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7081 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7141 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7201 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7321 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7381 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     7441 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7501 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     7561 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7621 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     7681 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     7741 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aattt
//
