LOCUS       sequence-335553-        3639 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 3639)
  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 3639)
  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-SLC25A3_STOP
FEATURES             Location/Qualifiers
     source          1..3639
                     /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(1196..1295)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1312..1331)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1313..1331)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1336..1352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1336..1352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1454..1473)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1465..2274
                     /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(1532..1551)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2421..3009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2910..2929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3163..3180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3339..3366)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3458..3544)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3593..3615
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3607..3624
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3608..3624
                     /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 ccaccatgtt cagctccgtg
      121 gcacacctgg caagggcaaa cccttttaat accccacacc tgcagctggt gcacgacgga
      181 ctgggcgatc tgcggtctag ctcccctgga ccaacaggac agccacggag acccagaaac
      241 ctggcagcag cagcagtgga ggagcagtac tcctgtgact atggctctgg ccggttcttt
      301 atcctgtgcg gcctgggcgg catcatctcc tgtggaacca cacacaccgc cctggtgcca
      361 ctggacctgg tgaagtgcag aatgcaggtg gatcctcaga agtacaaggg catcttcaat
      421 ggcttttccg tgaccctgaa ggaggatggc gtgaggggac tggcaaaggg atgggcacct
      481 acattcctgg gctatagcat gcagggcctg tgcaagttcg gcttttacga ggtgtttaag
      541 gtgctgtata gcaacatgct gggcgaggag aatacctacc tgtggaggac atccctgtat
      601 ctggcagcat ccgcctctgc cgagttcttt gcagacatcg ccctggcacc aatggaggca
      661 gcaaaggtgc ggatccagac ccagcctggc tacgccaaca cactgagaga tgccgcccca
      721 aagatgtaca aggaggaggg cctgaaggcc ttctataagg gcgtggcccc cctgtggatg
      781 aggcagatcc cttacaccat gatgaagttc gcctgctttg agcgcacagt ggaggccctg
      841 tataagttcg tggtgcccaa gcctcggagc gagtgttcca agccagagca gctggtggtg
      901 accttcgtgg ccggctacat cgccggcgtg ttttgcgcaa tcgtgtccca cccagcagac
      961 tctgtggtga gcgtgctgaa taaggagaag ggctctagcg cctctctggt gctgaagagg
     1021 ctgggcttca agggcgtgtg gaagggcctg tttgcccgca tcatcatgat cggcaccctg
     1081 acagccctgc agtggttcat ctacgattct gtgaaggtgt acttccggct gccaaggcca
     1141 cctccaccag agatgcccga gagcctgaag aagaagctgg gcctgacaca gtaagaccca
     1201 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     1261 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     1321 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     1381 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     1441 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     1501 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1561 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     1621 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     1681 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1741 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     1801 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1861 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     1921 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     1981 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     2041 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     2101 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     2161 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     2221 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     2281 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     2341 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     2401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3061 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3121 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3181 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3241 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3301 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     3361 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     3421 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     3481 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     3541 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     3601 cgacgttgta aaacgacggc cagtcttaag ctcgggccc
//
