LOCUS       sequence-264522-        3738 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 3738)
  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 3738)
  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_SLC35A2
FEATURES             Location/Qualifiers
     source          1..3738
                     /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(1863..1962)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1979..1998)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1980..1998)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2003..2019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2003..2019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2121..2140)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2132..2941
                     /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(2199..2218)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3088..3676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3577..3596
                     /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 ccgtgggagc aggaggaagc accgcagcac caggccctgg
      721 cgccgtgtcc gccggcgccc tggagcctgg aacagcatct gccgcacacc ggagactgaa
      781 gtacatcagc ctggccgtgc tggtggtgca gaacgcctct ctgatcctga gcatcagata
      841 tgcccgcacc ctgccaggcg acaggttctt tgccaccaca gccgtggtca tggccgaggt
      901 gctgaagggc ctgacatgcc tgctgctgct gtttgcccag aagcgcggca atgtgaagca
      961 cctggtgctg ttcctgcacg aggccgtgct ggtgcagtac gtggataccc tgaagctggc
     1021 cgtgccatcc ctgatctaca cactgcagaa caatctgcag tatgtggcca tctctaacct
     1081 gcccgccgcc acctttcagg tgacatatca gctgaagatc ctgaccacag ccctgttcag
     1141 cgtgctgatg ctgaatcgga gcctgtccag actgcagtgg gcctccctgc tgctgctgtt
     1201 taccggcgtg gccatcgtgc aggcacagca ggcaggagga ggaggaccac ggcccctgga
     1261 ccagaacccc ggcgccggcc tggccgccgt ggtggcctct tgtctgagct ccggctttgc
     1321 cggcgtgtac ttcgagaaga tcctgaaggg ctctagcggc tccgtgtggc tgcggaatct
     1381 gcagctgggc ctgttcggaa ccgccctggg actggtggga ctgtggtggg ccgagggcac
     1441 cgccgtggcc acaagaggct tctttttcgg atacacacca gccgtgtggg gagtggtgct
     1501 gaaccaggcc tttggcggac tgctggtggc agtggtggtg aagtatgccg acaatatcct
     1561 gaagggcttc gccacctctc tgagcatcgt gctgtccaca gtggcctcta tcaggctgtt
     1621 tggcttccac gtggatcctc tgtttgcact gggagcagga ctggtcatcg gagccgtgta
     1681 cctgtatagc ctgccaaggg gagcagcaaa ggcaatcgcc tccgcctctg ccagcgcctc
     1741 cggcccttgc gtgcaccagc agccccctgg ccagccaccc cctccacagc tgtcctctca
     1801 cagaggcgat ctgatcaccg agcctttcct gccaaagctg ctgacaaagg tgaagggctc
     1861 cgacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1921 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     1981 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2041 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2101 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2161 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2221 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2281 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2341 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2401 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2461 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2521 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2581 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2641 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2701 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2761 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2821 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2881 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2941 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3001 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3061 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3121 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3181 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3241 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3301 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3361 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3421 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3481 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3541 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3601 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3661 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3721 gccttttgct cacatgtt
//
