LOCUS       sequence-369570-        3645 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 3645)
  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 3645)
  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-SLC25A28_STOP
FEATURES             Location/Qualifiers
     source          1..3645
                     /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(1202..1301)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1318..1337)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1319..1337)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1342..1358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1342..1358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1460..1479)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1471..2280
                     /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(1538..1557)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2427..3015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2916..2935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3169..3186
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3345..3372)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3464..3550)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3599..3621
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3613..3630
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3614..3630
                     /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 ccaccatgga gctggaggga
      121 aggggagcag gaggagtggc aggaggccct gccgccggcc caggccgctc tcccggcgag
      181 agcgccctgc tggacggctg gctgcagcgg ggcgtgggca gaggcgccgg cggcggcgag
      241 gccggcgcct gcaggccacc tgtgcgccag gaccctgata gcggcccaga ttacgaggcc
      301 ctgccagcag gagcaaccgt gaccacacac atggtggcag gagcagtggc aggaatcctg
      361 gagcactgcg tgatgtaccc tatcgactgc gtgaagacca ggatgcagtc cctgcagcca
      421 gatccagcag caaggtatcg caacgtgctg gaggccctgt ggcggatcat cagaacagag
      481 ggactgtgga ggccaatgag gggcctgaat gtgaccgcaa caggagcagg accagcacac
      541 gcactgtact tcgcctgcta tgagaagctg aagaagaccc tgtctgacgt gatccaccct
      601 ggcggcaaca gccacatcgc aaatggagca gcaggatgcg tggccacact gctgcacgat
      661 gccgccatga accctgccga ggtggtgaag cagcggatgc agatgtacaa ttccccatat
      721 cacagggtga ccgactgcgt gcgggccgtg tggcagaacg agggagcagg cgccttttac
      781 agatcttata ccacacagct gaccatgaat gtgccattcc aggccatcca ctttatgaca
      841 tacgagttcc tgcaggagca ctttaacccc cagcggagat ataatcctag ctcccacgtg
      901 ctgtctggag catgcgcagg agcagtggca gcagcagcca ccacacccct ggacgtgtgc
      961 aagaccctgc tgaacacaca ggagtccctg gccctgaatt ctcacatcac cggccacatc
     1021 acaggcatgg cctccgcctt ccggaccgtg taccaagtgg gaggagtgac agcctatttt
     1081 cggggcgtgc aggccagagt gatctaccag atccccagca ccgccatcgc ctggtccgtg
     1141 tacgagttct ttaagtatct gatcacaaag aggcaggagg agtggcgcgc cggcaagtaa
     1201 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     1261 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     1321 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     1381 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1441 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1501 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1561 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     1621 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     1681 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1741 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     1801 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1861 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     1921 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1981 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     2041 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     2101 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2161 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2221 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     2281 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     2341 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2401 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2461 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2521 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2581 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2641 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2701 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2761 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2821 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2881 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2941 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3001 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3061 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3121 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3181 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3241 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3301 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3361 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3421 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3481 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     3541 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     3601 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
