LOCUS       sequence-479332-        3963 bp    DNA     circular SYN 31-JUL-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 3963)
  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 3963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 31, 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-SLC36A3_STOP
FEATURES             Location/Qualifiers
     source          1..3963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(561..588)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(680..766)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     815..837
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     829..846
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     830..846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    862..961
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2381..2480)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2497..2516)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2498..2516)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2521..2537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2521..2537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2639..2658)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2650..3459
                     /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(2717..2736)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3606..3963,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
      541 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
      601 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
      661 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
      721 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
      781 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
      841 gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg
      901 ttgcaacaca ttgatgagca atgctttttt ataatgccaa ctttgtacaa aaaagcaggc
      961 tccaccatgt ccctgctggg cagggactac aactctgagc tgaatagcct ggataacggc
     1021 cctcagtccc catctgagag ctcctctagc atcacatctg agaatgtgca cccagcagga
     1081 gaggcaggac tgagcatgat gcagaccctg atccacctgc tgaagtgcaa tatcggaaca
     1141 ggactgctgg gactgccact ggccatcaag aacgcaggac tgctggtggg acccgtgagc
     1201 ctgctggcaa tcggcgtgct gaccgtgcac tgcatggtca tcctgctgaa ctgtgcccag
     1261 cacctgagcc agagactgca gaagaccttc gtgaattacg gcgaggccac aatgtatggc
     1321 ctggagacct gtcctaacac atggctgagg gcacacgccg tgtggggcag atacaccgtg
     1381 tccttcctgc tggtcatcac acagctgggc ttttgcagcg tgtacttcat gtttatggcc
     1441 gacaatctgc agcagatggt ggagaaggcc cacgtgacca gcaacatctg tcagccaagg
     1501 gagatcctga ccctgacacc catcctggat atccgctttt atatgctgat catcctgcct
     1561 ttcctgatcc tgctggtgtt tatccagaat ctgaaggtgc tgtccgtgtt ctctacactg
     1621 gccaacatca ccacactggg cagcatggcc ctgatctttg agtacatcat ggagggcatc
     1681 ccctatcctt ccaatctgcc actgatggcc aactggaaga ccttcctgct gttctttggc
     1741 accgccatct tcacatttga gggcgtgggc atggtgctgc ctctgaagaa tcagatgaag
     1801 cacccacagc agttcagctt tgtgctgtac ctgggcatgt ccatcgtgat catcctgtac
     1861 atcctgctgg gcacactggg ctatatgaag tttggctctg acacccaggc cagcatcaca
     1921 ctgaacctgc ccaattgctg gctgtaccag tccgtgaagc tgatgtattc tatcggcatc
     1981 ttctttacct atgccctgca gttccacgtg cccgccgaga tcatcatccc ttttgccatc
     2041 tcccaggtga gcgagtcctg ggccctgttc gtggacctga gcgtgcggtc cgccctggtg
     2101 tgcctgacct gcgtgagcgc catcctgatc ccaagactgg acctggtcat ctccctggtg
     2161 ggaagcgtga gctctagcgc cctggccctg atcatccccg ccctgctgga gatcgtgatc
     2221 ttctacagcg aggacatgtc ctgcgtgacc atcgccaagg atatcatgat ctccatcgtg
     2281 ggcctgctgg gctgtatctt tggcacatac caggccctgt atgagctgcc acagcccatc
     2341 tctcacagca tggccaacag caccggcgtg cacgcctaag acccagcttt cttgtacaaa
     2401 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     2461 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     2521 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2581 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2641 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     2701 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2761 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2821 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2881 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2941 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     3001 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     3061 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     3121 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     3181 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     3241 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     3301 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     3361 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     3421 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     3481 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     3541 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3601 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3661 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3721 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3781 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3841 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3901 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3961 acg
//
