LOCUS Exported 4302 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway entry clone with codon-optimized ORF sequence for subcloning into custom expression vectors. ACCESSION . VERSION . KEYWORDS pDONR221_SLC8B1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4302) TITLE RESOLUTE consortium SLC collection REFERENCE 2 (bases 1 to 4302) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4302 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(2427..2526) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2543..2562) /label=T7 /note="T7 promoter, forward primer" promoter complement(2544..2562) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2567..2583) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2567..2583) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2685..2704) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2696..3505 /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(2763..2782) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3652..4240 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4141..4160 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agcaggctcc accatggcag gccggagact gaacctgaga tgggccctgt ctgtgctgtg 721 cgtgctgctg atggcagaga ccgtgagcgg aacaagggga agctccaccg gagcacacat 781 ctctccccag tttcctgcca gcggcgtgaa tcagacaccc gtggtggact gcagaaaggt 841 gtgcggcctg aacgtgtccg acaggtgcga tttcatccgc accaatcctg actgtcactc 901 tgatggcggc tacctggatt atctggaggg catcttctgc cactttccac ctagcctgct 961 gcctctggca gtgaccctgt acgtgtcctg gctgctgtat ctgtttctga tcctgggcgt 1021 gacagccgcc aagttctttt gtccaaacct gtccgccatc tctaccacac tgaagctgag 1081 ccacaacgtg gccggcgtga ccttcctggc ctttggcaat ggcgccccag acatcttctc 1141 cgccctggtg gccttttctg atccacacac agcaggactg gccctgggcg ccctgttcgg 1201 agcaggcgtg ctggtgacca cagtggtggc aggaggaatc accatcctgc accctttcat 1261 ggcagcatcc aggccattct ttagagacat cgtgttttac atggtggccg tgttcctgac 1321 ctttctgatg ctgttcaggg gaagggtgac actggcatgg gcactgggat acctgggact 1381 gtacgtgttt tatgtggtga ccgtgatcct gtgcacatgg atctatcaga ggcagaggcg 1441 cggctctctg ttctgtccta tgccagtgac ccccgagatc ctgtccgact ctgaggagga 1501 tcgggtgtct agcaacacaa atagctacga ctatggcgat gagtacagac ctctgttctt 1561 ttatcaggag accacagccc agatcctggt gagggccctg aatccactgg actacatgaa 1621 gtggcggaga aagagcgcct attggaaggc cctgaaggtg ttcaagctgc ccgtggagtt 1681 tctgctgctg ctgaccgtcc cagtggtgga ccccgataag gacgatcaga actggaagcg 1741 ccccctgaat tgtctgcacc tggtcatctc ccctctggtc gtggtgctga ccctgcagtc 1801 tggcacatac ggcgtgtatg agatcggagg actggtgccc gtgtgggtgg tggtggtcat 1861 cgcaggaaca gccctggcat ccgtgacctt ctttgccaca agcgattccc agccaccaag 1921 gctgcactgg ctgttcgcct ttctgggctt tctgacctct gccctgtgga tcaacgccgc 1981 cgccacagag gtggtgaata tcctgaggtc tctgggcgtg gtgttccgcc tgagcaacac 2041 cgtgctggga ctgacactgc tggcatgggg caatagcatc ggcgacgcct tctccgactt 2101 caccctggca aggcagggat acccaagaat ggccttcagc gcctgctttg gcggcatcat 2161 cttcaacatc ctggtgggag tgggactggg atgtctgctg cagatctcta gaagccacac 2221 agaggtgaag ctggagccag atggcctgct ggtgtgggtg ctggcaggcg ccctgggact 2281 gagcctggtg tttagcctgg tgtccgtgcc cctgcagtgc ttccagctgt ccagggtgta 2341 cggcttttgt ctgctgctgt tctatctgaa ttttctggtg gtggccctgc tgaccgagtt 2401 cggcgtgatc cacctgaagt ccatggaccc agctttcttg tacaaagttg gcattataag 2461 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat 2521 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc 2581 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat 2641 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag 2701 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata 2761 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta 2821 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga 2881 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat 2941 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa 3001 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct 3061 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga 3121 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag 3181 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa 3241 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct 3301 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga 3361 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca 3421 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca 3481 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag 3541 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta 3601 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 3661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 3721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 3781 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc 3841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 3901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 3961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 4021 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 4081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 4141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 4201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 4261 tttacggttc ctggcctttt gctggccttt tgctcacatg tt //