LOCUS       sequence-405322-        4695 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 4695)
  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 4695)
  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-SLC12A8_STOP
FEATURES             Location/Qualifiers
     source          1..4695
                     /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(2252..2351)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2368..2387)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2369..2387)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2392..2408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2392..2408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2510..2529)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2521..3330
                     /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(2588..2607)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3477..4065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3966..3985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4219..4236
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4395..4422)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4514..4600)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4649..4671
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4663..4680
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4664..4680
                     /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 ccaccatgac ccagatgtcc
      121 caggtgcagg agctgttcca cgaggcagca cagcaggacg cactggcaca gccacagccc
      181 tggtggaaga cacagctgtt catgtgggag cccgtgctgt ttggcacctg ggatggcgtg
      241 ttcacaagct gcatgatcaa catcttcggc gtggtgctgt ttctgagaac cggctggctg
      301 gtgggcaata caggcgtgct gctgggcatg ttcctggtgt cctttgtgat cctggtggcc
      361 ctggtgaccg tgctgtctgg catcggcgtg ggagagagga gctccatcgg aagcggaggc
      421 gtgtactcca tgatctctag cgtgctggga ggacagaccg gaggcacaat cggcctgctg
      481 tacgtgtttg gccagtgcgt ggccggcgcc atgtatatca caggcttcgc cgagagcatc
      541 tccgacctgc tgggactggg aaacatctgg gccgtgcgcg gaatctccgt ggccgtgctg
      601 ctggccctgc tgggcatcaa tctggccggc gtgaagtgga tcatccggct gcagctgctg
      661 ctgctgtttc tgctggccgt gagcaccctg gacttcgtgg tgggctcctt tacacacctg
      721 gatccagagc acggcttcat cggctacagc cccgagctgc tgcagaacaa taccctgcct
      781 gactattccc caggcgagtc tttctttacc gtgtttggcg tgttctttcc agcagcaaca
      841 ggcgtgatgg caggcttcaa catgggaggc gatctgaggg agccagcagc atctatccca
      901 ctgggaagcc tggcagcagt gggaatctcc tggtttctgt acatcatctt cgtgtttctg
      961 ctgggagcaa tctgcacaag ggaggccctg agatatgact tcctgatcgc cgagaaggtg
     1021 tctctgatgg gcttcctgtt tctgctgggc ctgtacatct cctctctggc cagctgtatg
     1081 ggaggactgt atggagcacc aaggatcctg cagtgcatcg cacaggagaa agtgatcccc
     1141 gccctggcat gtctgggaca gggcaagggc cccaataaga cccctgtggc agcaatctgc
     1201 ctgaccagcc tggtgacaat ggccttcgtg tttgtgggcc aggtgaacgt gctggcccct
     1261 atcgtgacca tcaacttcat gctgacatac gtggccgtgg attactccta tttctctctg
     1321 agcatgtgct cctgttctct gacacctgtg ccagagcccg tgctgagaga gggagcagag
     1381 ggactgcact gttctgagca cctgctgctg gagaaggcac ctagctatgg atccgaggga
     1441 ccagcacagc gggtgctgga gggcaccctg ctggagttta caaaggacat ggatcagctg
     1501 ctgcagctga ccagaaagct ggagagctcc cagcctaggc agggagaggg aaacaggaca
     1561 ccagagtccc agaagaggaa gtctaagaag gccaccaagc agacactgca ggacagcttt
     1621 ctgctggatc tgaagtctcc ccctagcttc ccagtggaga tctccgacag gctgcctgca
     1681 gcaagctggg agggccagga gtcctgctgg aacaagcaga ccagcaagtc cgagggaacc
     1741 cagccagagg gaacatacgg agagcagctg gtgcctgagc tgtgcaatca gtctgagtct
     1801 agcggcgagg acttctttct gaagagcagg ctgcaggagc aggacgtgtg gcggagatct
     1861 accagctttt acacacacat gtgcaatcct tgggtgtctc tgctgggagc agtgggaagc
     1921 ctgctgatca tgttcgtgat ccagtgggtg tataccctgg tgaacatggg cgtggccgcc
     1981 atcgtgtact tttatatcgg cagagcatcc ccaggactgc acctgggatc cgcctctaat
     2041 ttctccttct ttcggtggat gagatctctg ctgctgccat cctgtaggtc tctgcgcagc
     2101 ccccaggagc agatcatcct ggcccctagc ctggccaagg tggatatgga gatgacccag
     2161 ctgacacagg agaacgccga cttcgccacc agggatcgct accaccactc ctctctggtg
     2221 aatagggagc agctgatgcc ccactattaa gacccagctt tcttgtacaa agttggcatt
     2281 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     2341 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     2401 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2461 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2521 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2581 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2641 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2701 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2761 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2821 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2881 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2941 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     3001 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     3061 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     3121 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     3181 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     3241 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3301 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3361 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3421 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3481 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3541 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3601 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3661 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3721 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3781 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3841 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3901 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3961 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4021 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4081 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4141 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4201 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4261 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4321 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     4381 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     4441 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     4501 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     4561 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     4621 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     4681 cttaagctcg ggccc
//
