LOCUS       sequence-260976-        3642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3642)
  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 3642)
  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
FEATURES             Location/Qualifiers
     source          1..3642
                     /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(1767..1866)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1883..1902)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1884..1902)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1907..1923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1907..1923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2025..2044)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2036..2845
                     /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(2103..2122)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2992..3580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3481..3500
                     /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 accatggagc tggagggaag gggagcagga ggagtggcag gaggccctgc
      721 cgccggccca ggccgctctc ccggcgagag cgccctgctg gacggctggc tgcagcgggg
      781 cgtgggcaga ggcgccggcg gcggcgaggc cggcgcctgc aggccacctg tgcgccagga
      841 ccctgatagc ggcccagatt acgaggccct gccagcagga gcaaccgtga ccacacacat
      901 ggtggcagga gcagtggcag gaatcctgga gcactgcgtg atgtacccta tcgactgcgt
      961 gaagaccagg atgcagtccc tgcagccaga tccagcagca aggtatcgca acgtgctgga
     1021 ggccctgtgg cggatcatca gaacagaggg actgtggagg ccaatgaggg gcctgaatgt
     1081 gaccgcaaca ggagcaggac cagcacacgc actgtacttc gcctgctatg agaagctgaa
     1141 gaagaccctg tctgacgtga tccaccctgg cggcaacagc cacatcgcaa atggagcagc
     1201 aggatgcgtg gccacactgc tgcacgatgc cgccatgaac cctgccgagg tggtgaagca
     1261 gcggatgcag atgtacaatt ccccatatca cagggtgacc gactgcgtgc gggccgtgtg
     1321 gcagaacgag ggagcaggcg ccttttacag atcttatacc acacagctga ccatgaatgt
     1381 gccattccag gccatccact ttatgacata cgagttcctg caggagcact ttaaccccca
     1441 gcggagatat aatcctagct cccacgtgct gtctggagca tgcgcaggag cagtggcagc
     1501 agcagccacc acacccctgg acgtgtgcaa gaccctgctg aacacacagg agtccctggc
     1561 cctgaattct cacatcaccg gccacatcac aggcatggcc tccgccttcc ggaccgtgta
     1621 ccaagtggga ggagtgacag cctattttcg gggcgtgcag gccagagtga tctaccagat
     1681 ccccagcacc gccatcgcct ggtccgtgta cgagttcttt aagtatctga tcacaaagag
     1741 gcaggaggag tggcgcgccg gcaaggaccc agctttcttg tacaaagttg gcattataag
     1801 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     1861 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1921 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1981 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2041 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2101 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2161 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2221 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2281 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2341 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2401 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2461 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2521 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2581 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2641 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2701 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2761 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2821 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2881 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2941 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3001 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3061 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3121 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3181 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3241 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3301 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3361 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3421 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3481 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3541 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3601 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
