LOCUS       sequence-479318-        4233 bp    DNA     circular SYN 01-AUG-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 4233)
  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 4233)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 1, 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-SLC49A3_STOP
FEATURES             Location/Qualifiers
     source          1..4233
                     /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(2651..2750)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2767..2786)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2768..2786)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2791..2807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2791..2807)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2909..2928)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2920..3729
                     /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(2987..3006)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3876..4233,1..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 tccaccatgg ccggccctac cgaggccgag acaggactgg cagagccacg ggccctgtgc
     1021 gcacagaggg gacacaggac ctacgcacgg agatgggtgt tcctgctggc catctccctg
     1081 ctgaactgtt ctaatgccac actgtggctg tcttttgccc cagtggcaga cgtgatcgca
     1141 gaggatctgg tgctgagcat ggagcagatc aactggctgt ccctggtgta tctggtggtg
     1201 tctacccctt tcggcgtggc cgccatctgg atcctggact ccgtgggact gagggcagca
     1261 acaatcctgg gagcctggct gaactttgcc ggctctgtgc tgcgcatggt gccttgcatg
     1321 gtggtgggca cccagaatcc attcgccttt ctgatgggag gacagagcct gtgcgcactg
     1381 gcacagagcc tggtcatctt cagcccagca aagctggccg ccctgtggtt tcctgagcac
     1441 cagagagcca ccgccaacat gctggccaca atgtccaatc cactgggcgt gctggtggcc
     1501 aacgtgctgt ctcccgtgct ggtgaagaag ggcgaggaca tcccactgat gctgggcgtg
     1561 tacaccatcc cagcaggagt ggtgtgcctg ctgagcacca tctgtctgtg ggagtccgtg
     1621 ccacctacac caccatctgc cggagcagcc agctccacct ccgagaagtt cctggatggc
     1681 ctgaagctgc agctgatgtg gaataaggcc tatgtgatcc tggccgtgtg cctgggagga
     1741 atgatcggca tctctgccag cttctccgcc ctgctggagc agatcctgtg cgccagcggc
     1801 cactctagcg gcttttccgg actgtgcggc gccctgttca tcacctttgg catcctgggc
     1861 gccctggccc tgggaccata cgtggacagg acaaagcact tcaccgaggc cacaaagatc
     1921 ggcctgtgcc tgttctctct ggcctgcgtg ccctttgccc tggtgagcca gctgcaggga
     1981 cagaccctgg ccctggcagc aacatgctct ctgctgggcc tgttcggctt tagcgtggga
     2041 ccagtggcaa tggagctggc agtggagtgt agcttccctg tgggagaggg agcagcaacc
     2101 ggcatgatct ttgtgctggg ccaggccgag ggcatcctga tcatgctggc catgaccgcc
     2161 ctgacagtga ggcgctccga gccttctctg agcacatgtc agcagggcga ggacccactg
     2221 gattggaccg tgagcctgct gctgatggca ggactgtgca cattcttttc ctgtatcctg
     2281 gccgtgttct ttcacacccc ttatcggaga ctgcaggccg agtctggcga gcctccaagc
     2341 acaaggaatg cagtgggagg agcagacagc ggaccaggag tggatcgggg cggcgccggc
     2401 agagccggcg tgctgggacc atccaccgca acacctgagt gcaccgcaag gggagccagc
     2461 ctggaggatc caagaggccc cggctccccc caccctgcct gtcacagggc aaccccaagg
     2521 gcacagggac cagcagccac agacgcacct agcaggccag gcagactggc aggcagggtg
     2581 caggcatccc gcttcatcga tccagcagga tctcactcct cttttagctc cccttgggtc
     2641 atcacataag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     2701 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     2761 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2821 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2881 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2941 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     3001 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     3061 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     3121 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     3181 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3241 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     3301 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     3361 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3421 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3481 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3541 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3601 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3661 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3721 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     3781 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3841 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3961 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4021 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4081 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4141 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4201 ggttcgtgca cacagcccag cttggagcga acg
//
