LOCUS Exported 7800 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses FLAG-tagged ZnT1-WT in mammalian cells. ACCESSION . VERSION . KEYWORDS FLAG-ZnT1-WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7800) 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 7800) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7800 /organism="synthetic DNA construct" /mol_type="other DNA" 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 2559..3181 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2758..2777) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(3205..3225) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3223..3520 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 3452..3471 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3574..4368 /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(3628..3647) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4238..4257 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5020..5154 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5057..5076) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5111..5130 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5159..5178) /label=T7 /note="T7 promoter, forward primer" promoter complement(5160..5178) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5192..5208) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5192..5208) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5205..5227) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5216..5232 /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(5240..5270) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5285..5306 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5423..5440) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5594..6182) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5674..5693) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6353..7213) /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 6976..6995 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7214..7318) /gene="bla" /label=AmpR promoter rep_origin 7345..7800 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7432..7451) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7642..7663 /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 cggccgcgat ggggtgttgg 1021 ggtcggaacc ggggccggct gctgtgcatg ctggcgctga ccttcatgtt catggtgctg 1081 gaggtggtgg tgagccgggt gacctcgtcg ctggcgatgc tctccgactc cttccacatg 1141 ctgtcggacg tgctggcgct ggtggtggcg ctggtggccg agcgcttcgc ccggcggacc 1201 cacgccaccc agaagaacac gttcggctgg atccgagccg aggtaatggg ggctctggtg 1261 aacgccatct tcctgactgg cctctgtttc gccatcctgc tggaggccat cgagcgcttc 1321 atcgagccgc acgagatgca gcagccgctg gtggtccttg gggtcggcgt ggccgggctg 1381 ctggtcaacg tgctggggct ctgcctcttc caccatcaca gcggcttcag ccaggactcc 1441 ggccacggcc actcgcacgg gggtcacggc cacggccacg gcctccccaa ggggcctcgc 1501 gttaagagca cccgccccgg gagcagcgac atcaacgtgg ccccgggcga gcagggtccc 1561 gaccaggagg agaccaacac cctggtggcc aataccagca actccaacgg gctgaaattg 1621 gaccccgcag acccagaaaa ccccagaagt ggtgatacag tggaagtaca agtgaatgga 1681 aatcttgtca gagaacctga ccatatggaa ctggaagaag atagggctgg acaacttaac 1741 atgcgtggag tttttctgca tgtccttgga gatgccttgg gttcagtgat tgtagtagta 1801 aatgccttag tcttttactt ttcttggaaa ggttgttctg aaggggattt ttgtgtgaat 1861 ccatgtttcc ctgacccctg caaagcattt gtagaaataa ttaatagtac tcatgcatca 1921 gtttatgagg ctggtccttg ctgggtgcta tatttagatc caactctttg tgttgtaatg 1981 gtttgtatac ttctttacac aacctatcca ttacttaagg aatctgctct tattcttcta 2041 caaactgttc ctaaacaaat tgatatcaga aatttgataa aagaacttcg aaatgttgaa 2101 ggagttgagg aagttcatga attacatgtt tggcaacttg ctggaagcag aatcattgcc 2161 actgctcaca taaaatgtga agatccaaca tcatacatgg aggtggctaa aaccattaaa 2221 gacgtttttc ataatcacgg aattcacgct actaccattc agcctgaatt tgctagtgta 2281 ggctctaaat caagtgtagt tccgtgtgaa cttgcctgca gaacccagtg tgctttgaag 2341 caatgttgtg ggacactacc acaagcccct tctggaaagg atgcagaaaa gaccccagca 2401 gttagcattt cttgtttaga acttagtaac aatctagaga agaagcccag gaggactaaa 2461 gctgaaaaca tccctgctgt tgtgatagag attaaaaaca tgccaaacaa acaacctgaa 2521 tcatctttgt agggtaccag tcgactctag aggatcccgg gtggcatccc tgtgacccct 2581 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa 2641 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga 2701 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct 2761 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct 2821 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga 2881 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc 2941 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat 3001 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc 3061 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga 3121 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat 3181 tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3361 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 3481 cctctgagct attccagaag tagtgaggag gcttttttgg aggaattgat cagcttggga 3541 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 3601 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3661 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 3721 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 3781 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 3841 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3901 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3961 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 4021 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 4081 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 4141 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 4201 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 4261 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 4321 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 4381 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4441 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 4501 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat cccctcgcga 4561 gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg cagtgcaaat 4621 ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc gaactgcagg 4681 agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg cgcctgatac 4741 agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc cgttttccgc ctgaggtcac 4801 tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag ctgctcacca cccactcgcc 4861 aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gccgatcata 4921 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc 4981 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat 5041 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 5101 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaattcc 5161 ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt 5221 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5281 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5401 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5461 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5521 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5581 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5641 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5701 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5761 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 5821 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5881 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5941 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6001 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6061 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6121 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 6181 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6241 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 6301 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 6361 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 6421 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6481 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6541 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6601 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6661 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6721 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6781 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6841 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6901 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6961 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 7021 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 7081 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 7141 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 7201 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7261 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7321 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg 7381 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 7441 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 7501 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 7561 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 7621 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 7681 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa 7741 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt //