LOCUS       sequence-262965-        4845 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 4845)
  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 4845)
  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_SLC30A5
FEATURES             Location/Qualifiers
     source          1..4845
                     /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(2970..3069)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3086..3105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3087..3105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3110..3126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3110..3126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3228..3247)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3239..4048
                     /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(3306..3325)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4195..4783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4684..4703
                     /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 accatggagg agaagtacgg cggcgacgtg ctggcaggcc caggaggagg
      721 aggaggactg ggccctgtgg atgtgccatc cgcccggctg accaagtaca tcgtgctgct
      781 gtgcttcaca aagtttctga aggccgtggg cctgttcgag tcttatgacc tgctgaaggc
      841 cgtgcacatc gtgcagttca tctttatcct gaagctgggc accgccttct ttatggtgct
      901 gttccagaag ccttttagct ccggcaagac catcacaaag caccagtgga tcaagatctt
      961 taagcacgcc gtggccggct gcatcatctc tctgctgtgg ttctttggac tgaccctgtg
     1021 cggaccactg cggacactgc tgctgttcga gcacagcgat atcgtggtca tctccctgct
     1081 gtctgtgctg tttacctcta gcggaggagg accagcaaag acaagaggag ccgccttctt
     1141 tatcatcgcc gtgatctgcc tgctgctgtt cgacaacgac gatctgatgg ccaagatggc
     1201 cgagcaccct gagggccacc acgattccgc cctgacccac atgctgtata cagcaatcgc
     1261 cttcctggga gtggcagacc acaagggagg cgtgctgctg ctggtgctgg ccctgtgctg
     1321 taaagtgggc tttcacaccg cctcccggaa gctgtctgtg gatgtgggag gagcaaagag
     1381 actgcaggcc ctgagccacc tggtgtccgt gctgctgctg tgcccctggg tcatcgtgct
     1441 gtccgtgacc acagagtcta aggtggagag ctggttctcc ctgatcatgc cttttgccac
     1501 agtgatcttc tttgtgatga tcctggactt ctacgtggat tctatctgca gcgtgaagat
     1561 ggaggtgagc aagtgtgccc ggtatggctc tttcccaatc tttatcagcg ccctgctgtt
     1621 cggcaacttt tggacccacc ccatcacaga ccagctgaga gccatgaata aggcagcaca
     1681 ccaggagtcc accgagcacg tgctgtctgg aggagtggtg gtgagcgcca tcttctttat
     1741 cctgagcgcc aacatcctgt cctctccttc caagcggggc cagaagggca ccctgatcgg
     1801 atactcccca gagggcacac ctctgtataa tttcatgggc gacgcctttc agcacagctc
     1861 ccagtctatc ccaaggttca tcaaggagag cctgaagcag atcctggagg agtccgattc
     1921 tcgccagatc ttctactttc tgtgcctgaa cctgctgttc acctttgtgg agctgtttta
     1981 tggcgtgctg acaaatagcc tgggcctgat ctccgacggc ttccacatgc tgtttgattg
     2041 tagcgccctg gtcatgggac tgttcgccgc cctgatgtcc aggtggaagg caaccagaat
     2101 cttttcttac ggctatggca ggatcgagat cctgtctggc ttcatcaacg gcctgtttct
     2161 gatcgtgatc gccttcttcg tgttcatgga gagcgtggca aggctgatcg acccacctga
     2221 gctggatacc cacatgctga caccagtgtc cgtgggcggc ctgatcgtga atctgatcgg
     2281 catctgcgcc ttttctcacg cacacagcca cgcacacgga gcatcccagg gctcttgtca
     2341 ctctagcgac cacagccact cccaccacat gcacggacac tccgatcacg gacacggaca
     2401 cagccacgga tccgccggag gaggcatgaa cgccaatatg aggggcgtgt tcctgcacgt
     2461 gctggcagac accctgggat ccatcggcgt gatcgtgtct acagtgctga tcgagcagtt
     2521 cggctggttt atcgccgacc ctctgtgcag cctgttcatc gccatcctga tctttctgtc
     2581 cgtggtgcca ctgatcaagg atgcctgtca ggtgctgctg ctgaggctgc cacccgagta
     2641 cgagaaggag ctgcacatcg ccctggagaa gatccagaag atcgagggcc tgatcagcta
     2701 tagggatccc cacttctgga ggcactctgc cagcatcgtg gcaggcacca tccacatcca
     2761 ggtgacatcc gacgtgctgg agcagcgcat cgtgcagcag gtgaccggca tcctgaagga
     2821 tgccggcgtg aacaatctga caatccaggt ggagaaggag gcctacttcc agcacatgtc
     2881 tggcctgagc accggctttc acgacgtgct ggccatgaca aagcagatgg agagcatgaa
     2941 gtactgtaag gatggcacct atatcatgga cccagctttc ttgtacaaag ttggcattat
     3001 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     3061 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     3121 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     3181 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     3241 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     3301 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     3361 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     3421 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     3481 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     3541 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3601 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3661 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3721 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3781 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3841 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3901 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3961 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     4021 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     4081 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     4141 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4321 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
