LOCUS       sequence-262699-        4263 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 4263)
  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 4263)
  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_SLC40A1
FEATURES             Location/Qualifiers
     source          1..4263
                     /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(2388..2487)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2504..2523)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2505..2523)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2528..2544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2528..2544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2646..2665)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2657..3466
                     /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(2724..2743)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3613..4201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4102..4121
                     /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 gggcaggcga ccacaatagg cagagaggat gctgtggatc
      721 cctggcagat tacctgacat ctgccaagtt tctgctgtat ctgggacaca gcctgtccac
      781 ctggggcgac agaatgtggc acttcgccgt gtccgtgttt ctggtggagc tgtacggcaa
      841 ctctctgctg ctgacagccg tgtatggact ggtggtggca ggatccgtgc tggtgctggg
      901 agcaatcatc ggcgactggg tggataagaa tgccaggctg aaggtggccc agaccagcct
      961 ggtggtgcag aacgtgtccg tgatcctgtg cggcatcatc ctgatgatgg tgttcctgca
     1021 caagcacgag ctgctgacca tgtaccacgg ctgggtgctg acatcttgtt atatcctgat
     1081 catcaccatc gccaacatcg ccaatctggc cagcaccgcc acagccatca caatccagag
     1141 ggactggatc gtggtggtgg caggagagga tcgctccaag ctggccaaca tgaatgccac
     1201 aatccggaga atcgatcagc tgaccaatat cctggcccct atggccgtgg gccagatcat
     1261 gacctttggc tctccagtga tcggctgcgg cttcatctct ggctggaacc tggtgagcat
     1321 gtgcgtggag tacgtgctgc tgtggaaggt gtatcagaag acacccgccc tggcagtgaa
     1381 ggcaggcctg aaggaggagg agaccgagct gaagcagctg aatctgcaca aggacacaga
     1441 gcccaagcct ctggagggca cccacctgat gggcgtgaag gattctaaca tccacgagct
     1501 ggagcacgag caggagccaa catgcgcaag ccagatggca gagcccttcc ggacctttag
     1561 agacggctgg gtgtcctact ataatcagcc cgtgttcctg gcaggaatgg gactggcctt
     1621 tctgtacatg acagtgctgg gcttcgattg catcaccaca ggctacgcct atacccaggg
     1681 cctgtctggc agcatcctgt ccatcctgat gggcgcctct gccatcacag gcatcatggg
     1741 caccgtggcc tttacatggc tgaggcgcaa gtgtggcctg gtgagaaccg gactgatcag
     1801 cggactggca cagctgtcct gcctgatcct gtgcgtgatc agcgtgttca tgcctggcag
     1861 cccactggac ctgtccgtgt ctccattcga ggatatcagg tctcgcttta tccagggcga
     1921 gagcatcacc cccacaaaga tccctgagat caccacagag atctacatgt ccaacggcag
     1981 caattccgcc aacatcgtgc cagagaccag cccagagtcc gtgcctatca tctctgtgag
     2041 cctgctgttt gcaggcgtga tcgcagcaag gatcggactg tggagcttcg acctgaccgt
     2101 gacacagctg ctgcaggaga atgtgatcga gagcgagcgc ggcatcatca acggcgtgca
     2161 gaattccatg aactacctgc tggatctgct gcactttatc atggtcatcc tggccccaaa
     2221 ccccgaggcc tttggcctgc tggtgctgat ctccgtgtct ttcgtggcca tgggccacat
     2281 catgtatttc cggtttgccc agaatacact gggcaacaag ctgttcgcat gtggaccaga
     2341 cgcaaaggag gtgagaaagg agaatcaggc caacaccagc gtggtggacc cagctttctt
     2401 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2461 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2521 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2581 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2641 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2701 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2761 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2821 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2881 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2941 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3001 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3061 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3121 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3181 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3241 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3301 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3361 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3421 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3481 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3541 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3721 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4261 gtt
//
