LOCUS       sequence-356918-        4296 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 4296)
  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 4296)
  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-SLC15A3_STOP
FEATURES             Location/Qualifiers
     source          1..4296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1853..1952)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1969..1988)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1970..1988)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1993..2009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1993..2009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2111..2130)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2122..2931
                     /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(2189..2208)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3078..3666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3567..3586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3820..3837
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3996..4023)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4115..4201)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4250..4272
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4264..4281
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4265..4281
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgcc agcccccagg
      121 gcccgcgagc agccaagggt gcccggcgag cgccagcctc tgctgccaag gggcgcccgc
      181 ggcccacgga gatggaggag ggcagcagga gcagccgtgc tgctggtgga gatgctggag
      241 agggcagcct tctttggcgt gacagccaac ctggtgctgt acctgaattc tacaaatttc
      301 aattggacag gcgaacaggc aaccagagcc gccctggtgt ttctgggagc aagctatctg
      361 ctggcacctg tgggaggatg gctggcagac gtgtacctgg gccggtatag agccgtggcc
      421 ctgtctctgc tgctgtacct ggcagcaagc ggactgctgc cagcaaccgc cttccctgac
      481 ggcaggagct ccttttgcgg cgagatgcca gcatcccctc tgggaccagc atgcccatct
      541 gccggatgtc ctcgctctag ccctagccca tactgtgccc cagtgctgta tgcaggactg
      601 ctgctgctgg gactggcagc atcctctgtg cggtctaacc tgacaagctt cggcgccgac
      661 caggtcatgg atctgggcag agacgccacc cggagattct ttaactggtt ttactggagc
      721 atcaatctgg gagccgtgct gtccctgctg gtggtggcct tcatccagca gaatatctcc
      781 tttctgctgg gctattctat cccagtgggc tgcgtgggcc tggccttctt tatcttcctg
      841 tttgccacac ccgtgttcat caccaagccc cctatgggct cccaggtgag ctccatgctg
      901 aagctggccc tgcagaactg ctgtccccag ctgtggcaga ggcactctgc cagggatagg
      961 cagtgtgcac gggtgctggc agacgagaga agcccacagc ctggagcatc ccctcaggag
     1021 gatatcgcca attttcaggt gctggtgaag atcctgccag tgatggtgac actggtgccc
     1081 tactggatgg tgtatttcca gatgcagagc acctacgtgc tgcagggcct gcacctgcac
     1141 atccctaaca tctttcctgc caacccagcc aatatctccg tggccctgag ggcacaggga
     1201 tctagctata caatcccaga ggcctggctg ctgctggcca atgtggtggt ggtgctgatc
     1261 ctggtgcccc tgaaggatag gctgatcgac cctctgctgc tgaggtgcaa gctgctgcca
     1321 agcgccctgc agaagatggc actgggcatg ttctttggct tcacatccgt gatcgtggca
     1381 ggcgtgctgg agatggagcg gctgcactac atccaccaca acgagaccgt gtcccagcag
     1441 atcggcgagg tgctgtacaa tgccgcccca ctgtctatct ggtggcagat cccccagtat
     1501 ctgctgatcg gcatcagcga gatcttcgcc tccatccccg gcctggagtt tgcctattct
     1561 gaggccccta gaagcatgca gggcgccatc atgggcatct tcttttgtct gtccggagtg
     1621 ggatctctgc tgggatcctc tctggtggcc ctgctgtccc tgcctggagg atggctgcac
     1681 tgcccaaagg atttcggcaa catcaacaat tgtaggatgg acctgtactt ctttctgctg
     1741 gcaggaatcc aggcagtgac cgccctgctg ttcgtgtgga tcgcaggccg gtatgagaga
     1801 gcatctcagg gaccagcaag ccactcccgc tttagccggg acagaggcta agacccagct
     1861 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1921 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1981 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     2041 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     2101 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     2161 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     2221 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2281 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2341 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     2401 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     2461 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2521 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2581 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2641 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     2701 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2761 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2821 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2881 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2941 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     3001 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     3061 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3121 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3181 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3601 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3661 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3721 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3781 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3841 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3901 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3961 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     4021 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     4081 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     4141 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     4201 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     4261 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
