LOCUS       sequence-430413-        3498 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 3498)
  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 3498)
  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-SLC25A32_STOP
FEATURES             Location/Qualifiers
     source          1..3498
                     /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(1055..1154)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1171..1190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1172..1190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1195..1211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1195..1211)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1313..1332)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1324..2133
                     /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(1391..1410)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2280..2868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2769..2788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3022..3039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3198..3225)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3317..3403)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3452..3474
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3466..3483
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3467..3483
                     /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 cggacaggga
      121 cagtctgcca gcggaagctc cgcctggtct acagtgttta ggcacgtgcg ctacgagaac
      181 ctgatcgcag gcgtgtccgg aggcgtgctg tctaatctgg ccctgcaccc actggacctg
      241 gtgaagatcc ggttcgccgt gagcgatggc ctggagctga gacccaagta caacggcatc
      301 ctgcactgcc tgaccacaat ctggaagctg gacggactga ggggactgta tcagggagtg
      361 accccaaaca tctggggagc aggactgtct tggggactgt acttcttttt ctataatgcc
      421 atcaagagct acaagaccga gggaagggca gagagactgg aggccacaga gtatctggtg
      481 agcgccgcag aggcaggagc aatgaccctg tgcatcacaa accctctgtg ggtgaccaag
      541 acaaggctga tgctgcagta cgacgccgtg gtgaattccc cacaccgcca gtataagggc
      601 atgtttgata ccctggtgaa gatctacaag tatgagggcg tgaggggcct gtacaagggc
      661 tttgtgccag gactgttcgg cacatcccac ggcgccctgc agttcatggc ctacgagctg
      721 ctgaagctga agtataacca gcacatcaat aggctgccag aggcacagct gtctaccgtg
      781 gagtatatca gcgtggccgc cctgtccaag atctttgccg tggccgccac atacccttat
      841 caggtggtgc gggccagact gcaggaccag cacatgtttt actccggcgt gatcgatgtg
      901 atcaccaaga catggcggaa ggagggcgtg ggcggcttct ataagggcat cgcccccaac
      961 ctgatcagag tgacccctgc ctgctgtatc acatttgtgg tgtacgagaa cgtgagccac
     1021 ttcctgctgg atctgaggga gaagcgcaag taagacccag ctttcttgta caaagttggc
     1081 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1141 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1201 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1261 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1321 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1381 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1441 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1501 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1561 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1621 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1681 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1741 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1801 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1861 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1921 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1981 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2041 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2101 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2161 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2221 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2281 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2341 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2401 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2461 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2521 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2581 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2641 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2701 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2761 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2821 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2881 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2941 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3001 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3061 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3121 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3181 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3241 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3301 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3361 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3421 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3481 agtcttaagc tcgggccc
//
