LOCUS       sequence-260431-        4269 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 4269)
  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 4269)
  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_SLC41A2
FEATURES             Location/Qualifiers
     source          1..4269
                     /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(2394..2493)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2510..2529)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2511..2529)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2534..2550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2534..2550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2652..2671)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2663..3472
                     /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(2730..2749)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3619..4207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4108..4127
                     /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 accatgacca acagcaaggg ccggtccatc accgataaga caagcggagg
      721 acctagctcc ggaggaggat tcgtggactg gaccctgaga ctgaacacaa tccagagcga
      781 taagtttctg aatctgctgc tgtccatggt gccagtgatc tatcagaaga accaggagga
      841 caggcacaag aaggccaatg gcatctggca ggatggcctg tccaccgccg tgcagacatt
      901 cagcaatcgc tccgagcagc acatggagta ccactctttc agcgagcagt cttttcacgc
      961 caacaatggc cacgcctcta gctcctgcag ccagaagtat gacgattacg ccaactataa
     1021 ttactgtgat ggcagggaga ccagcgagac cacagccatg ctgcaggacg aggatatctc
     1081 tagcgacggc gatgaggacg ccatcgtgga ggtgacaccc aagctgccta aggagtcctc
     1141 tggcatcatg gccctgcaga tcctggtgcc tttcctgctg gcaggatttg gaaccgtgtc
     1201 cgccggaatg gtgctggaca tcgtgcagca ctgggaggtg ttccggaagg tgacagaggt
     1261 gtttatcctg gtgccagccc tgctgggcct gaagggcaac ctggagatga ccctggcctc
     1321 tagactgagc acagccgtga atatcggcaa gatggatagc cccatcgaga agtggaacct
     1381 gatcatcggc aatctggccc tgaagcaggt gcaggccacc gtggtgggct tcctggcagc
     1441 agtggcagca atcatcctgg gctggatccc cgagggcaag tactatctgg accactccat
     1501 cctgctgtgc agctcctctg tggccacagc ctttatcgcc tccctgctgc agggcatcat
     1561 catggtgggc gtgatcgtgg gctctaagaa gaccggcatc aaccctgaca atgtggccac
     1621 accaatcgcc gcctctttcg gcgatctgat caccctggcc atcctggcct ggatctccca
     1681 gggcctgtat tcttgtctgg agacctacta ttacatcagc cccctggtgg gcgtgttctt
     1741 tctggccctg acacctatct ggatcatcat cgcagcaaag cacccagcaa ccaggacagt
     1801 gctgcacagc ggatgggagc ctgtgatcac cgccatggtc atcagctcca tcggcggcct
     1861 gatcctggac accacagtgt ccgatcccaa cctggtgggc atcgtggtgt atacccctgt
     1921 gatcaacggc atcggcggca atctggtggc catccaggcc tccaggatct ctacatacct
     1981 gcacctgcac tccatcccag gagagctgcc agacgagcca aagggatgct attacccatt
     2041 ccgcaccttc tttggccccg gcgtgaacaa taagtctgcc caggtgctgc tgctgctggt
     2101 catccccggc cacctgatct ttctgtatac aatccacctg atgaagtctg gccacaccag
     2161 cctgacaatc atcttcatcg tggtgtacct gtttggcgcc gtgctgcagg tgtttaccct
     2221 gctgtggatc gccgactgga tggtgcacca cttctggcgg aagggcaagg atcccgactc
     2281 cttttctatc ccttacctga ccgccctggg cgatctgctg ggaacagccc tgctggccct
     2341 gagcttccac tttctgtggc tgatcggcga tagagacggc gatgtgggcg acgacccagc
     2401 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2461 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2521 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2581 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2641 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2701 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2761 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2821 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2881 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2941 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3001 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3061 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3121 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3181 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3241 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3301 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3361 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3421 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3481 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3541 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3601 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3661 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3721 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3781 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3841 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3901 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3961 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4021 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4081 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4141 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4201 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4261 tcacatgtt
//
