LOCUS       sequence-262940-        3999 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 3999)
  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 3999)
  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_SLC36A2
FEATURES             Location/Qualifiers
     source          1..3999
                     /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(2124..2223)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2240..2259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2241..2259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2264..2280)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2264..2280)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2382..2401)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2393..3202
                     /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(2460..2479)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3349..3937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3838..3857
                     /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 accatgagcg tgaccaagtc cacagaggga ccacagggag cagtggccat
      721 caagctggac ctgatgtctc cccctgagag cgccaagaag ctggagaata aggactccac
      781 cttcctggat gagagcccct ccgagtctgc cggcctgaag aagaccaagg gcatcacagt
      841 gtttcaggcc ctgatccacc tggtgaaggg caatatggga acaggaatcc tgggactgcc
      901 tctggcagtg aagaacgcag gcatcctgat gggcccactg tctctgctgg tcatgggctt
      961 catcgcctgc cactgtatgc acatcctggt gaagtgcgcc cagaggttct gtaagcgcct
     1021 gaataagcct tttatggact acggcgatac cgtgatgcac ggcctggagg ccaacccaaa
     1081 tgcctggctg cagaaccacg cacactgggg ccggcacatc gtgagcttct ttctgatcat
     1141 cacacagctg ggcttctgct gcgtgtacat cgtgtttctg gccgacaacc tgaagcaggt
     1201 ggtggaggcc gtgaattcca ccacaaacaa ttgctactct aacgagaccg tgatcctgac
     1261 cccaacaatg gattctaggc tgtatatgct gagcttcctg ccctttctgg tgctgctggt
     1321 gctgatccgg aatctgagaa tcctgaccat cttctccatg ctggccaaca tctccatgct
     1381 ggtgtctctg gtcatcatca tccagtacat cacacaggag atccctgacc caagcaggct
     1441 gccactggtg gcatcctgga agacctatcc cctgttcttt ggcacagcca tcttcagctt
     1501 tgagtccatc ggcgtggtgc tgcccctgga gaacaagatg aagaatgccc ggcacttccc
     1561 tgccatcctg agcctgggca tgtccatcgt gacctctctg tacatcggca tggccgccct
     1621 gggctatctg agatttggcg acgatatcaa ggccagcatc tccctgaacc tgcccaattg
     1681 ctggctgtac cagagcgtga agctgctgta tatcgccggc atcctgtgca catacgccct
     1741 gcagttctat gtgcccgccg agatcatcat cccttttgcc atctctcggg tgagcaccag
     1801 atgggcactg ccactggacc tgtctatcag gctggtcatg gtgtgcctga catgtctgct
     1861 ggccatcctg atccctcgcc tggacctggt catcagcctg gtgggatctg tgagcggaac
     1921 cgccctggcc ctgatcatcc caccactgct ggaggtgacc acattctaca gcgagggcat
     1981 gtcccctctg accatcttta aggatgccct gatctccatc ctgggcttcg tgggctttgt
     2041 ggtgggcaca tatcaggccc tggacgagct gctgaagtcc gaggattctc acccattctc
     2101 caactctacc acatttgtga gagacccagc tttcttgtac aaagttggca ttataagaaa
     2161 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2221 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2281 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2341 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2401 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2461 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2521 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2581 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2641 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2701 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2761 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2821 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2881 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2941 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3001 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3061 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3121 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3181 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3241 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3301 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3361 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3421 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3481 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3541 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3601 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3661 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3721 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3781 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3841 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3901 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3961 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
