LOCUS       sequence-260974-        3453 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 3453)
  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 3453)
  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_SLC25A2
FEATURES             Location/Qualifiers
     source          1..3453
                     /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(1578..1677)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1694..1713)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1695..1713)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1718..1734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1718..1734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1836..1855)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1847..2656
                     /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(1914..1933)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2803..3391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3292..3311
                     /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 accatgaagt ccggaccagg aatccaggca gcaatcgacc tgaccgcagg
      721 agcagcagga ggaaccgcat gcgtgctgac aggacagccc ttcgatacca tcaaggtgaa
      781 gatgcagaca tttcctgacc tgtacaaggg cctgaccgat tgtttcctga agacatacgc
      841 ccaagtgggc ctgaggggct tttataaggg caccggaccc gccctgatgg catacgtggc
      901 agagaacagc gtgctgttca tgtgctatgg cttctgtcag cagtttgtgc gcaaggtggc
      961 cggcatggac aagcaggcca agctgtccga tctgcagaca gcagcagcag gatccttcgc
     1021 ctctgccttt gccgccctgg ccctgtgccc taccgagctg gtgaagtgtc ggctgcagac
     1081 aatgtacgag atggagatgt ctggcaagat cgccaagagc cacaatacca tctggtccgt
     1141 ggtgaagggc atcctgaaga aggacggccc actgggcttt tatcacggcc tgagctccac
     1201 actgctgcag gaggtgcccg gctacttctt tttctttggc ggctatgagc tgtctcggag
     1261 cttctttgcc agcggcagat ccaaggacga gctgggccct gtgcacctga tgctgtctgg
     1321 aggagtggca ggaatctgcc tgtggctggt ggtgttccca gtggattgta tcaagtctcg
     1381 gatccaggtg ctgagcatgt acggcaagca ggccggcttt atcggcaccc tgctgtccgt
     1441 ggtgagaaac gagggcatcg tggccctgta ttctggcctg aaggccacaa tgatcagggc
     1501 aatcccagca aatggcgccc tgttcgtggc ctacgagtat agccgcaaga tgatgatgaa
     1561 gcagctggag gcctatgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1621 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1681 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1741 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1801 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1861 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1921 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     1981 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2041 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2101 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2161 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2221 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2281 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2341 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2401 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2461 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2521 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2581 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2641 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2701 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2761 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2821 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2881 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2941 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3001 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3061 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3121 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3181 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3241 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3301 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3361 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3421 cctggccttt tgctggcctt ttgctcacat gtt
//
