LOCUS       sequence-253735-        4230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4230)
  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 4230)
  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_SLC49A3
FEATURES             Location/Qualifiers
     source          1..4230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2355..2454)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2471..2490)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2472..2490)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2495..2511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2495..2511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2613..2632)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2624..3433
                     /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(2691..2710)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3580..4168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4069..4088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggccg gccctaccga ggccgagaca ggactggcag agccacgggc
      721 cctgtgcgca cagaggggac acaggaccta cgcacggaga tgggtgttcc tgctggccat
      781 ctccctgctg aactgttcta atgccacact gtggctgtct tttgccccag tggcagacgt
      841 gatcgcagag gatctggtgc tgagcatgga gcagatcaac tggctgtccc tggtgtatct
      901 ggtggtgtct acccctttcg gcgtggccgc catctggatc ctggactccg tgggactgag
      961 ggcagcaaca atcctgggag cctggctgaa ctttgccggc tctgtgctgc gcatggtgcc
     1021 ttgcatggtg gtgggcaccc agaatccatt cgcctttctg atgggaggac agagcctgtg
     1081 cgcactggca cagagcctgg tcatcttcag cccagcaaag ctggccgccc tgtggtttcc
     1141 tgagcaccag agagccaccg ccaacatgct ggccacaatg tccaatccac tgggcgtgct
     1201 ggtggccaac gtgctgtctc ccgtgctggt gaagaagggc gaggacatcc cactgatgct
     1261 gggcgtgtac accatcccag caggagtggt gtgcctgctg agcaccatct gtctgtggga
     1321 gtccgtgcca cctacaccac catctgccgg agcagccagc tccacctccg agaagttcct
     1381 ggatggcctg aagctgcagc tgatgtggaa taaggcctat gtgatcctgg ccgtgtgcct
     1441 gggaggaatg atcggcatct ctgccagctt ctccgccctg ctggagcaga tcctgtgcgc
     1501 cagcggccac tctagcggct tttccggact gtgcggcgcc ctgttcatca cctttggcat
     1561 cctgggcgcc ctggccctgg gaccatacgt ggacaggaca aagcacttca ccgaggccac
     1621 aaagatcggc ctgtgcctgt tctctctggc ctgcgtgccc tttgccctgg tgagccagct
     1681 gcagggacag accctggccc tggcagcaac atgctctctg ctgggcctgt tcggctttag
     1741 cgtgggacca gtggcaatgg agctggcagt ggagtgtagc ttccctgtgg gagagggagc
     1801 agcaaccggc atgatctttg tgctgggcca ggccgagggc atcctgatca tgctggccat
     1861 gaccgccctg acagtgaggc gctccgagcc ttctctgagc acatgtcagc agggcgagga
     1921 cccactggat tggaccgtga gcctgctgct gatggcagga ctgtgcacat tcttttcctg
     1981 tatcctggcc gtgttctttc acacccctta tcggagactg caggccgagt ctggcgagcc
     2041 tccaagcaca aggaatgcag tgggaggagc agacagcgga ccaggagtgg atcggggcgg
     2101 cgccggcaga gccggcgtgc tgggaccatc caccgcaaca cctgagtgca ccgcaagggg
     2161 agccagcctg gaggatccaa gaggccccgg ctccccccac cctgcctgtc acagggcaac
     2221 cccaagggca cagggaccag cagccacaga cgcacctagc aggccaggca gactggcagg
     2281 cagggtgcag gcatcccgct tcatcgatcc agcaggatct cactcctctt ttagctcccc
     2341 ttgggtcatc acagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2401 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2461 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2521 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaataatat catcatgaac
     2581 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2641 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2701 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2761 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2821 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2881 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2941 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3001 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3061 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3121 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3181 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3241 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3301 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3361 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3421 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3481 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3721 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4201 ggccttttgc tggccttttg ctcacatgtt
//
