LOCUS       sequence-382439-        4194 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 4194)
  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 4194)
  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-SLC38A4_STOP
FEATURES             Location/Qualifiers
     source          1..4194
                     /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(1751..1850)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1867..1886)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1868..1886)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1891..1907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1891..1907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2009..2028)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2020..2829
                     /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(2087..2106)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2976..3564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3465..3484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3718..3735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3894..3921)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4013..4099)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4148..4170
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4162..4179
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4163..4179
                     /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 ccctatggag
      121 ctgaggaacg tgaatatcga gccagacgat gagagctcct ctggagagtc cgccccagac
      181 tcttacatcg gcatcggcaa ttctgagaag gccgccatga gctcccagtt cgccaacgag
      241 gataccgaga gccagaagtt cctgacaaat ggctttctgg gcaagaagaa gctggccgac
      301 tacgccgatg agcaccaccc tggcaccaca agcttcggca tgtctagctt taacctgtcc
      361 aatgccatca tgggctctgg catcctgggc ctgagctatg ccatggccaa caccggcatc
      421 atcctgttca tcatcatgct gctggccgtg gccatcctgt ccctgtactc tgtgcacctg
      481 ctgctgaaga cagccaagga gggcggctcc ctgatctatg agaagctggg agagaaggcc
      541 ttcggctggc ctggcaagat cggagccttc gtgagcatca ccatgcagaa catcggcgcc
      601 atgtcctctt acctgttcat catcaagtat gagctgccag aagtgatccg ggcctttatg
      661 ggcctggagg agaatacagg cgagtggtac ctgaacggca attatctgat catcttcgtg
      721 agcgtgggca tcatcctgcc tctgtccctg ctgaagaacc tgggctacct gggctatacc
      781 agcggcttct ccctgacatg catggtgttc tttgtgagcg tggtcatcta caagaagttt
      841 cagatcccat gtcccctgcc tgtgctggac cactctgtgg gcaacctgag cttcaacaat
      901 accctgccaa tgcacgtggt catgctgccc aacaattccg agagctccga cgtgaacttc
      961 atgatggatt acacacacag aaaccccgcc ggcctggacg agaatcaggc caagggcagc
     1021 ctgcacgatt ccggcgtgga gtatgaggcc cactccgacg ataagtgcga gcccaagtac
     1081 ttcgtgttta attctaggac cgcctatgcc atccctatcc tggtgttcgc cttcgtgtgc
     1141 caccctgagg tgctgccaat ctactccgag ctgaaggatc gctctcggag aaagatgcag
     1201 accgtgtcta acatcagcat cacaggcatg ctggtcatgt atctgctggc cgccctgttc
     1261 ggctacctga ccttttatgg agaggtggag gacgagctgc tgcacgcata cagcaaggtg
     1321 tatacactgg atatcccact gctgatggtg cggctggccg tgctggtggc agtgaccctg
     1381 acagtgccca tcgtgctgtt ccctatcaga acctctgtga tcacactgct gttcccaaag
     1441 aggcccttta gctggatccg ccactttctg atcgccgccg tgctgatcgc cctgaacaac
     1501 gtgctggtca tcctggtgcc caccatcaag tacatcttcg gctttatcgg cgcctctagc
     1561 gccacaatgc tgatcttcat cctgccagcc gtgttttatc tgaagctggt gaagaaggag
     1621 accttccggt ccccacagaa agtgggcgcc ctgatctttc tggtggtggg catcttcttt
     1681 atgatcggca gcatggccct gatcatcatc gactggatct acgatccccc taactccaag
     1741 caccactaag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1801 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1861 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     1921 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1981 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2041 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2101 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2161 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2221 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2281 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     2341 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2401 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2461 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2521 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2581 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2641 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2701 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2761 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2821 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     2881 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     2941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3121 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3721 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3781 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3841 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     3901 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     3961 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     4021 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     4081 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     4141 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccc
//
