LOCUS       sequence-416478-        3858 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3858)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 3858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221-SLC38A8_STOP
FEATURES             Location/Qualifiers
     source          1..3858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(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(1983..2082)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2099..2118)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2100..2118)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2123..2139)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2123..2139)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2241..2260)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2252..3061
                     /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(2319..2338)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3208..3796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3697..3716
                     /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 accatggagg gacagacccc aggatccagg ggactgcctg agaagccaca
      721 ccccgccacc gccgccgcca cactgagctc catgggcgcc gtgttcatcc tgatgaagtc
      781 cgccctggga gcaggactgc tgaacttccc ttgggccttt tctaaggcag gaggagtggt
      841 gccagccttc ctggtggagc tggtgtccct ggtgtttctg atctctggac tggtcatcct
      901 gggatacgca gcagccgtgt ctggacaggc cacatatcag ggagtggtgc ggggactgtg
      961 cggaccagcc atcggcaagc tgtgcgaggc ctgtttcctg ctgaatctgc tgatgatcag
     1021 cgtggccttt ctgagagtga tcggcgacca gctggagaag ctgtgcgatt ctctgctgag
     1081 cggaacccca cctgcacctc agccatggta cgcagaccag cggttcacac tgcctctgct
     1141 gagcgtgctg gtcatcctgc cactgtctgc ccccagagag atcgcctttc agaagtacac
     1201 cagcatcctg ggcacactgg cagcatgcta cctggccctg gtcatcaccg tgcagtacta
     1261 tctgtggcct cagggcctgg tgagagagtc ccacccttcc ctgtctccag cctcctggac
     1321 cagcgtgttc agcgtgttcc caacaatctg cttcggcttt cagtgtcacg aggccgccgt
     1381 gagcatctat tgttccatga ggaagcgcag cctgtcccac tgggcactgg tgtccgtgct
     1441 gtctctgctg gcctgctgtc tgatctactc cctgaccggc gtgtatggct tcctgacctt
     1501 tggcacagag gtgagcgccg acgtgctgat gtcctaccca ggcaacgata tggtcatcat
     1561 cgtggccagg gtgctgttcg ccgtgagcat cgtgacagtg tatcccatcg tgctgttcct
     1621 gggccgcagc gtgatgcagg acttttggcg gagatcctgc ctgggaggat ggggaccatc
     1681 tgccctggca gatccaagcg gactgtgggt gaggatgcca ctgaccatcc tgtgggtgac
     1741 cgtgacactg gccatggccc tgtttatgcc tgatctgtct gagatcgtga gcatcatcgg
     1801 cggcatctct agcttcttta tcttcatctt tcccggcctg tgcctgatct gtgcaatggg
     1861 agtggagcca atcggaccta gggtgaagtg ctgtctggaa gtgtggggcg tggtgtctgt
     1921 gctggtgggc accttcatct ttggacagag cacagcagca gccgtgtggg agatgttcta
     1981 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2041 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2101 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2161 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2221 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2281 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2341 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2401 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2461 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2521 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2581 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2641 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2701 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2761 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2821 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2881 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2941 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3001 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3061 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3121 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3181 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3241 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3301 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3361 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3421 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3481 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3541 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3601 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3661 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3721 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3781 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3841 gccttttgct cacatgtt
//
