LOCUS       sequence-479340-        4056 bp    DNA     circular SYN 15-DEC-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 4056)
  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 4056)
  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-SLC2A5_STOP
FEATURES             Location/Qualifiers
     source          1..4056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(541..568)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(660..746)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     795..817
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     809..826
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     810..826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    842..941
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2454..2553)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2570..2589)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2571..2589)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2594..2610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2594..2610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2712..2731)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2723..3532
                     /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(2790..2809)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3679..4056,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
      541 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
      601 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
      661 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      721 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      781 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
      841 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaca ttgatgagca
      901 atgctttttt ataatgccaa ctttgtacaa aaaagcaggc tccaccatgg agcagcagga
      961 ccagagcatg aaggagggcc ggctgaccct ggtgctggcc ctggccacac tgatcgcagc
     1021 cttcggcagc tcatttcaat acgggtacaa tgtcgcggcc gtgaattccc ctgccctgct
     1081 gatgcagcag ttctataacg agacctacta tggccggaca ggcgagttca tggaggattt
     1141 tccactgacc ctgctgtgga gcgtgacagt gtccatgttc ccatttggcg gcttcatcgg
     1201 atccctgctg gtgggaccac tggtgaataa gtttggcaga aagggcgccc tgctgttcaa
     1261 caatatcttt tctatcgtgc ctgccatcct gatgggctgc tctagggtgg ccaccagctt
     1321 tgagctgatc atcatcagca ggctgctggt gggaatctgt gcaggcgtgt ctagcaacgt
     1381 ggtgccaatg tatctgggcg agctggcacc aaagaatctg aggggcgccc tgggagtggt
     1441 gccacagctg ttcatcaccg tgggcatcct ggtggcccag atctttggcc tgcgcaacct
     1501 gctggccaat gtggacggat ggccaatcct gctgggactg acaggagtgc ctgccgccct
     1561 gcagctgctg ctgctgccat tctttcccga gagccctcgg tacctgctga tccagaagaa
     1621 ggacgaggca gcagccaaga aggccctgca gaccctgaga ggatgggact ccgtggatag
     1681 ggaggtggca gagatcagac aggaggatga ggcagagaag gcagcaggct tcatctctgt
     1741 gctgaagctg tttaggatgc gcagcctgag gtggcagctg ctgtccatca tcgtgctgat
     1801 gggaggacag cagctgtctg gagtgaacgc catctactat tacgccgacc agatctatct
     1861 gagcgccgga gtgccagagg agcacgtgca gtacgtgacc gcaggaacag gagcagtgaa
     1921 cgtggtcatg accttctgcg ccgtgtttgt ggtggagctg ctgggccgga gactgctgct
     1981 gctgctgggc ttctctatct gtctgatcgc atgctgcgtg ctgaccgccg ccctggccct
     2041 gcaggataca gtgagctgga tgccatatat ctccatcgtg tgcgtgatct cttacgtgat
     2101 cggacacgca ctgggaccaa gcccaatccc tgccctgctg atcaccgaga tcttcctgca
     2161 gtcctctagg ccatccgcct ttatggtggg aggatccgtg cactggctgt ctaacttcac
     2221 agtgggcctg atcttccctt ttatccagga gggcctgggc ccatatagct tcatcgtgtt
     2281 tgccgtgatc tgtctgctga ccacaatcta catctttctg atcgtgcccg agaccaaggc
     2341 caagacattc atcgagatca accagatctt taccaagatg aataaggtgt ccgaggtgta
     2401 ccctgagaag gaggagctga aggagctgcc ccctgtgaca tctgagcagt aagacccagc
     2461 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2521 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2581 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2641 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2701 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2761 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2821 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2881 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2941 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3001 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3061 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3121 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3181 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3241 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3301 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3361 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3421 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3481 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3541 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3601 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3661 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3721 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3781 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3841 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3901 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3961 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4021 cagcttggag cgaacgacct acaccgaact gagata
//
