LOCUS       sequence-408930-        5295 bp    DNA     circular SYN 15-DEC-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 5295)
  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 5295)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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-SLC12A9_STOP
FEATURES             Location/Qualifiers
     source          1..5295
                     /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(2852..2951)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2968..2987)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2969..2987)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2992..3008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2992..3008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3110..3129)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3121..3930
                     /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(3188..3207)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4077..4665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4566..4585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4819..4836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4995..5022)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5114..5200)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5249..5271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5263..5280
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5264..5280
                     /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 ccaccatggc cagcgagagc
      121 tcccctctgc tggcatacag gctgctggga gaggagggcg tggccctgcc tgccaatgga
      181 gcaggcggac caggaggagc atctgccaga aagctgagca cctttctggg cgtggtggtg
      241 ccaacagtgc tgtctatgtt cagcatcgtg gtgtttctgc ggatcggctt cgtggtggga
      301 cacgcaggac tgctgcaggc cctggcaatg ctgctggtgg cctatttcat cctggccctg
      361 accgtgctgt ccgtgtgcgc catcgcaaca aatggagcag tgcagggagg aggcgcctac
      421 tttatgatca gcagaaccct gggacctgaa gtgggaggat ccatcggcct gatgttctat
      481 ctggccaacg tgtgcggatg tgccgtgagc ctgctgggac tggtggagtc cgtgctggac
      541 gtgtttggag cagatgcaac aggaccatcc ggactgaggg tgctgcctca gggatacggc
      601 tggaatctgc tgtatggctc tctgctgctg ggcctggtgg gaggcgtgtg caccctggga
      661 gcaggcctgt acgcccgcgc cagcttcctg acatttctgc tggtgtctgg cagcctggcc
      721 agcgtgctga tctccttcgt ggcagtggga ccacgggata tcagactgac ccctaggcca
      781 ggaccaaacg gatctagcct gccccctaga ttcggccact ttaccggctt caactcctct
      841 acactgaagg acaatctggg cgccggctac gccgaggatt ataccacagg cgccgtgatg
      901 aactttgcct ccgtgttcgc cgtgctgttt aatggctgca caggcatcat ggccggcgcc
      961 aacatgtctg gcgagctgaa ggaccctagc agggcaatcc cactgggaac catcgtggca
     1021 gtggcctaca ccttcttcgt gtacgtgctg ctgttctttc tgagctcctt tacctgtgac
     1081 aggacactgc tgcaggagga ttacggcttc tttcgcgcca tctctctgtg gccacccctg
     1141 gtgctgatcg gcatctatgc caccgccctg tccgcctcta tgtctagcct gatcggagca
     1201 agcaggatcc tgcacgcact ggcaagagac gatctgttcg gcgtgatcct ggcacctgca
     1261 aaggtggtgt ccaggggagg aaatccatgg gcagccgtgc tgtactcttg gggactggtg
     1321 cagctggtgc tgctggcagg caagctgaac accctggcag ccgtggtgac agtgttttac
     1381 ctggtggcct atgccgccgt ggatctgtcc tgcctgtctc tggagtgggc aagcgcccca
     1441 aatttcaggc ccacctttag cctgttctcc tggcacacat gcctgctggg cgtggcctcc
     1501 tgtctgctga tgatgttcct gatctcccca ggagcagcag gaggatctct gctgctgatg
     1561 ggactgctgg ccgccctgct gaccgcaagg ggaggacctt cctcttgggg ctacgtgagc
     1621 caggccctgc tgtttcacca ggtgcggaag tatctgctga ggctggacgt gcgcaaggat
     1681 cacgtgaagt tctggagacc ccagctgctg ctgctggtgg gcaatccaag gggcgccctg
     1741 cctctgctga ggctggccaa ccagctgaag aagggaggac tgtacgtgct gggacacgtg
     1801 accctgggcg acctggattc tctgcctagc gatccagtgc agccccagta tggagcatgg
     1861 ctgagcctgg tggacagggc acaggtgaag gcctttgtgg atctgacact gagcccatcc
     1921 gtgcggcagg gagcacagca cctgctgaga atctccggac tgggaggaat gaagccaaac
     1981 accctggtgc tgggcttcta cgacgatgcc cctccacagg accactttct gacagatcca
     2041 gccttctccg agccagcaga ctctaccagg gagggcagct cccctgccct gtctacactg
     2101 ttcccacctc caagggcacc tggaagccca cgcgccctga atccacagga ctatgtggcc
     2161 accgtggccg atgccctgaa gatgaacaag aatgtggtgc tggcaagagc atctggcgcc
     2221 ctgccacctg agaggctgtc caggggatct ggaggcacaa gccagctgca ccacgtggac
     2281 gtgtggccac tgaacctgct gaggcctaga ggaggaccag gatacgtgga cgtgtgcgga
     2341 ctgtttctgc tgcagatggc aaccatcctg ggaatggtgc cagcatggca ctccgccagg
     2401 ctgcgcatct tcctgtgcct gggaccaaga gaggcacctg gagcagcaga gggccggctg
     2461 agagccctgc tgtctcagct gaggatccgc gccgaggtgc aggaggtggt gtggggagag
     2521 ggagcaggag caggagagcc agaggccgag gaggagggcg actttgtgaa tagcggcagg
     2581 ggcgatgccg aggccgaggc cctggcccgc tccgccaacg ccctggtgcg ggcccagcag
     2641 ggcagaggca ccggcggagg accaggagga cctgagggag gcgacgccga gggccccatc
     2701 accgccctga cattcctgta tctgcctagg ccaccagcag acccagcaag gtaccccaga
     2761 tatctggccc tgctggagac cctgacacgc gatctgggac caaccctgct ggtgcacgga
     2821 gtgacacccg tgacctgtac agacctgtaa gacccagctt tcttgtacaa agttggcatt
     2881 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     2941 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     3001 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     3061 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     3121 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     3181 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     3241 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     3301 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     3361 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     3421 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     3481 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     3541 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     3601 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     3661 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     3721 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     3781 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     3841 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3901 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3961 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     4021 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4081 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4141 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4201 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4261 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4321 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4381 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4441 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4501 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4561 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4621 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4681 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4741 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4801 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4861 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4921 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     4981 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     5041 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     5101 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     5161 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     5221 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     5281 cttaagctcg ggccc
//
