LOCUS       sequence-265774-        4122 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 4122)
  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 4122)
  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_SLC2A2
FEATURES             Location/Qualifiers
     source          1..4122
                     /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(2247..2346)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2363..2382)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2364..2382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2387..2403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2387..2403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2505..2524)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2516..3325
                     /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(2583..2602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3472..4060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3961..3980
                     /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 accatgaccg aggacaaggt gaccggcaca ctggtgttca ccgtgatcac
      721 agccgtgctg ggctctttcc agtttggcta cgatatcggc gtgatcaatg ccccccagca
      781 ggtcatcatc agccactata ggcacgtgct gggcgtgcct ctggacgatc gcaaggccat
      841 caacaattac gtgatcaact ccaccgacga gctgcccaca atcagctatt ccatgaatcc
      901 caagcctacc ccatgggcag aggaggagac agtggcagca gcacagctga tcaccatgct
      961 gtggtctctg agcgtgagct ccttcgccgt gggaggaatg acagccagct tctttggagg
     1021 atggctgggc gataccctgg gaaggatcaa ggccatgctg gtggccaata tcctgtccct
     1081 ggtgggcgcc ctgctgatgg gcttttccaa gctgggccct tctcacatcc tgatcatcgc
     1141 cggcaggtcc atctctggcc tgtactgcgg cctgatctcc ggcctggtgc ctatgtatat
     1201 cggcgagatc gcaccaaccg ccctgagggg cgccctgggc acattccacc agctggccat
     1261 cgtgaccggc atcctgatct ctcagatcat cggcctggag tttatcctgg gcaactacga
     1321 cctgtggcac atcctgctgg gactgtctgg cgtgcgggca atcctgcaga gcctgctgct
     1381 gttcttttgt ccagagagcc ccagatacct gtatatcaag ctggacgagg aggtgaaggc
     1441 caagcagtcc ctgaagaggc tgcgcggcta cgacgatgtg acaaaggata tcaacgagat
     1501 gcggaaggag agagaggagg cctctagcga gcagaaggtg tccatcatcc agctgttcac
     1561 caattcctct tatcggcagc ctatcctggt ggccctgatg ctgcacgtgg cccagcagtt
     1621 ttccggcatc aacggcatct tctactatag cacctccatc tttcagacag ccggcatctc
     1681 taagccagtg tacgccacaa tcggcgtggg cgccgtgaat atggtgttca ccgccgtgag
     1741 cgtgtttctg gtggagaagg ccggccggag aagcctgttc ctgatcggca tgtccggcat
     1801 gttcgtgtgc gccatcttta tgtctgtggg cctggtgctg ctgaacaagt tttcctggat
     1861 gtcttatgtg agcatgatcg ccatcttcct gtttgtgagc ttctttgaga tcggccccgg
     1921 ccctatccca tggttcatgg tggcagagtt cttttcccag ggaccaaggc ctgccgccct
     1981 ggcaatcgcc gccttttcta actggacatg caatttcatc gtggccctgt gctttcagta
     2041 catcgccgat ttctgtggcc catacgtgtt cttcctgttc gccggcgtgc tgctggcctt
     2101 caccctgttt acattcttta aggtgcccga gaccaagggc aagagcttcg aggagatcgc
     2161 cgccgagttt cagaagaagt ctggaagcgc ccacagacca aaggcagcag tggagatgaa
     2221 gtttctgggc gccaccgaga cagtggaccc agctttcttg tacaaagttg gcattataag
     2281 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2341 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2401 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2461 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2521 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2581 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2641 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2701 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2761 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2821 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2881 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2941 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3001 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3061 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3121 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3181 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3241 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3301 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3361 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3421 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3601 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4081 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
