LOCUS       sequence-360556-        2991 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 2991)
  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 2991)
  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-SLC48A1_STOP
FEATURES             Location/Qualifiers
     source          1..2991
                     /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(548..647)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(664..683)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(665..683)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(688..704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(688..704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(806..825)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             817..1626
                     /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(884..903)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1773..2361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2262..2281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2515..2532
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2691..2718)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2810..2896)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2945..2967
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2959..2976
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2960..2976
                     /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 ccaccatggc cccctctagg
      121 ctgcagctgg gactgagggc agcatacagc ggcatcagct ccgtggccgg cttttccatc
      181 ttcctggtgt ggaccgtggt gtataggcag cctggaacag cagcaatggg aggactggca
      241 ggcgtgctgg ccctgtgggt gctggtgacc cacgtgatgt acatgcagga ctattggcgg
      301 acatggctga agggcctgag aggcttcttt ttcgtgggcg tgctgttttc cgccgtgtct
      361 atcgccgcct tttgcacctt cctggtgctg gccatcacaa gacaccagtc cctgaccgac
      421 cccacatctt actatctgtc tagcgtgtgg agctttatct ccttcaagtg ggccttcctg
      481 ctgtccctgt acgcccaccg gtatagagcc gactttgccg atatctctat cctgagcgat
      541 ttctaagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
      601 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
      661 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
      721 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
      781 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
      841 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
      901 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
      961 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1021 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1081 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1141 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1201 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     1261 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     1321 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     1381 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     1441 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     1501 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     1561 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     1621 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     1681 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     1741 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1801 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1861 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1921 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1981 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2041 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2101 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2161 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2221 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2281 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2341 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2401 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2461 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2521 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2581 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2641 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     2701 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     2761 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     2821 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     2881 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     2941 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc c
//
