LOCUS       sequence-265841-        3678 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 3678)
  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 3678)
  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_SLC30A7
FEATURES             Location/Qualifiers
     source          1..3678
                     /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"
     regulatory      1793..1802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1803..1902)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1919..1938)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1920..1938)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1943..1959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1943..1959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2061..2080)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2072..2881
                     /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(2139..2158)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3028..3616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3517..3536
                     /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 accatgctgc ccctgagcat caaggacgat gagtacaagc cccctaagtt
      721 taacctgttc ggcaagatct ctggctggtt ccggtccatc ctgtctgaca agaccagcag
      781 aaacctgttc tttttcctgt gcctgaatct gtcctttgcc ttcgtggagc tgctgtatgg
      841 catctggtct aattgtctgg gcctgatcag cgactccttt cacatgtttt tcgattctac
      901 agcaatcctg gcaggactgg cagcaagcgt gatctccaag tggcgggaca acgatgcctt
      961 tagctacggc tatgtgagag ccgaggtgct ggccggcttc gtgaatggcc tgtttctgat
     1021 cttcaccgcc tttttcatct tcagcgaggg agtggagagg gccctggcac cacccgatgt
     1081 gcaccacgag cgcctgctgc tggtgtccat cctgggcttt gtggtgaacc tgatcggcat
     1141 cttcgtgttc aagcacggag gacacggaca ctctcacgga agcggccatg gacactctca
     1201 cagcctgttc aatggcgccc tggaccaggc acacggacac gtggatcact gccactccca
     1261 cgaggtgaag cacggagcag cacactctca cgaccacgcc cacggccacg gccactttca
     1321 ctcccacgat ggcccctctc tgaaggagac cacaggccct agcaggcaga tcctgcaggg
     1381 cgtgttcctg cacatcctgg ccgacacact gggaagcatc ggcgtgatcg catccgccat
     1441 catgatgcag aactttggcc tgatgatcgc cgatccaatc tgttccatcc tgatcgccat
     1501 cctgatcgtg gtgtctgtga tccccctgct gagggagagc gtgggcatcc tgatgcagcg
     1561 cacccctcca ctgctggaga attctctgcc tcagtgctac cagagggtgc agcagctgca
     1621 gggcgtgtat agcctgcagg agcagcactt ctggaccctg tgctccgacg tgtacgtggg
     1681 cacactgaag ctgatcgtgg caccagacgc agatgcaagg tggatcctgt cccagaccca
     1741 caacatcttt acacaggccg gcgtgagaca gctgtatgtg cagatcgatt tcgccgccat
     1801 ggacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1861 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     1921 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1981 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2041 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2101 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2161 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2221 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2281 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2341 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2401 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2461 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2521 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2581 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2641 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2701 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2761 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2821 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2881 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2941 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3661 gccttttgct cacatgtt
//
