LOCUS Exported 4335 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway entry clone with codon-optimized ORF sequence for subcloning into custom expression vectors. ACCESSION . VERSION . KEYWORDS pDONR221_SLC13A1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4335) TITLE RESOLUTE consortium SLC collection REFERENCE 2 (bases 1 to 4335) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4335 /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" regulatory 1103..1112 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2460..2559) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2576..2595) /label=T7 /note="T7 promoter, forward primer" promoter complement(2577..2595) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2600..2616) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2600..2616) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2718..2737) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2729..3538 /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(2796..2815) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3685..4273 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4174..4193 /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 accatgaagt tctttagcta catcctggtg tatcggagat tcctgtttgt 721 ggtgtttaca gtgctggtgc tgctgccact gcccatcgtg ctgcacacaa aggaggccga 781 gtgcgcctac accctgtttg tggtggccac attctggctg accgaggccc tgccactgtc 841 tgtgaccgcc ctgctgccaa gcctgatgct gcctatgttc ggcatcatgc cttccaagaa 901 ggtggcctct gcctacttta aggacttcca cctgctgctg atcggcgtga tctgtctggc 961 cacctccatc gagaagtgga acctgcacaa gagaatcgcc ctgaagatgg tcatgatggt 1021 gggcgtgaat cctgcctggc tgacactggg ctttatgagc tccaccgcct tcctgtctat 1081 gtggctgtct aacaccagca cagccgccat ggtcatgcca atcgccgagg ccgtggtgca 1141 gcagatcatc aatgccgagg ccgaggtgga ggccacacag atgacctact tcaacggctc 1201 caccaatcac ggcctggaga tcgacgagtc tgtgaacggc cacgagatca atgagcggaa 1261 ggagaagaca aagcctgtgc caggctataa caatgatacc ggcaagatct ctagcaaggt 1321 ggagctggag aagaacagcg gcatgaggac aaagtaccgc accaagaagg gccacgtgac 1381 acggaagctg acctgcctgt gcatcgccta ttcctctacc atcggcggcc tgaccacaat 1441 caccggcaca tccaccaacc tgatctttgc cgagtacttc aatacacggt atcctgactg 1501 cagatgtctg aactttggca gctggttcac cttttccttc ccagccgccc tgatcatcct 1561 gctgctgtcc tggatctggc tgcagtggct gtttctgggc tttaatttca aggagatgtt 1621 caagtgcggc aagacaaaga ccgtgcagca gaaggcctgt gccgaagtga tcaagcagga 1681 gtaccagaag ctgggcccca tcaggtatca ggagatcgtg accctggtgc tgttcatcat 1741 catggccctg ctgtggtttt ctagggaccc aggattcgtg cctggatgga gcgccctgtt 1801 ttccgagtac cctggcttcg ccacagatag caccgtggcc ctgctgatcg gcctgctgtt 1861 ctttctgatc cccgccaaga cactgaccaa gaccacacct acaggcgaga tcgtggcctt 1921 tgattatagc ccactgatca cctggaagga gtttcagtcc ttcatgccat gggacatcgc 1981 aatcctggtg ggaggaggat tcgcactggc agatggatgc gaggagtccg gactgtctaa 2041 gtggatcggc aataagctgt ctccactggg aagcctgcca gcatggctga tcatcctgat 2101 cagctccctg atggtgacat ccctgaccga ggtggcctct aaccccgcca caatcaccct 2161 gttcctgcca atcctgagcc cactggcaga ggcaatccac gtgaatcctc tgtacatcct 2221 gatcccatct acactgtgca ccagctttgc cttcctgctg ccagtggcaa acccacctaa 2281 tgccatcgtg ttttcctatg gccacctgaa agtgatcgac atggtgaagg caggactggg 2341 agtgaacatc gtgggagtgg ccgtggtcat gctgggcatc tgtacatgga tcgtgccaat 2401 gttcgatctg tacacctatc ctagctgggc cccagccatg tccaatgaga ccatgcccga 2461 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa 2521 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta 2581 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa 2641 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc 2701 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc 2761 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt 2821 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt 2881 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa 2941 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga 3001 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata 3061 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc 3121 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca 3181 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg 3241 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt 3301 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg 3361 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg 3421 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat 3481 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc 3541 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc 3601 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc 3661 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 3721 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 3781 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 3841 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 3901 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 3961 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 4021 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 4081 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 4141 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 4201 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 4261 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 4321 ttttgctcac atgtt //