LOCUS       sequence-335512-        3477 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 3477)
  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 3477)
  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-SLC25A47_STOP
FEATURES             Location/Qualifiers
     source          1..3477
                     /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(1034..1133)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1150..1169)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1151..1169)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1174..1190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1174..1190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1292..1311)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1303..2112
                     /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(1370..1389)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2259..2847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2748..2767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3001..3018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3177..3204)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3296..3382)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3431..3453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3445..3462
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3446..3462
                     /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 cttcgtggca
      121 ggagcaatcg gaggcgtgtg cggcgtggcc gtgggctacc ccctggatac cgtgaaggtg
      181 aggatccaga ccgagcctaa gtatacaggc atctggcact gcgtgcggga cacataccac
      241 cgggagagag tgtggggctt ctatagagga ctgtccctgc ccgtgtgcac cgtgtccctg
      301 gtgagctccg tgtcttttgg cacataccgc cactgcctgg cccacatctg taggctgcgc
      361 tatggcaacc cagacgccaa gcccaccaag gccgatatca cactgagcgg atgtgcatcc
      421 ggactggtga gggtgtttct gacctctccc acagaggtgg ccaaggtgcg cctgcagacc
      481 cagacacagg cccagaagca gcagcggaga ctgtccgcct ctggacctct ggcagtgcca
      541 cctatgtgcc cagtgccacc agcatgtcct gagccaaagt acaggggacc tctgcactgc
      601 ctggcaaccg tggcaagaga ggagggactg tgcggactgt ataagggatc tagcgccctg
      661 gtgctgaggg acggacacag cttcgccaca tactttctgt cctatgccgt gctgtgcgag
      721 tggctgtctc ctgcaggaca cagcagacca gatgtgcctg gcgtgctggt ggcaggagga
      781 tgtgcaggcg tgctggcctg ggccgtggcc accccaatgg acgtgatcaa gtctaggctg
      841 caggccgatg gacagggaca gaggcgctac aggggactgc tgcactgcat ggtgacaagc
      901 gtgagggagg agggaccacg cgtgctgttc aagggcctgg tgctgaactg ctgtcgggcc
      961 ttccccgtga atatggtggt gtttgtggca tatgaggccg tgctgaggct ggcaagaggc
     1021 ctgctgacct aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1081 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1141 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1201 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     1261 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1321 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1381 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     1441 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1501 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1561 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     1621 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1681 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1741 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1801 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     1861 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1921 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     1981 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2041 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2101 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2161 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     2221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2401 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2881 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2941 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3001 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3061 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3121 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3181 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     3241 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     3301 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     3361 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     3421 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
