LOCUS       sequence-262937-        4491 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 4491)
  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 4491)
  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_SLC39A4
FEATURES             Location/Qualifiers
     source          1..4491
                     /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(2616..2715)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2732..2751)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2733..2751)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2756..2772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2756..2772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2874..2893)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2885..3694
                     /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(2952..2971)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3841..4429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4330..4349
                     /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 accatggcct ctctggtgag cctggagctg ggactgctgc tggccgtgct
      721 ggtggtgacc gcaacagcaa gcccacctgc aggactgctg tctctgctga caagcggaca
      781 gggcgccctg gatcaggagg ccctgggagg actgctgaac accctggcag acagggtgca
      841 ctgcgcaaat ggaccctgcg gcaagtgtct gtctgtggag gatgccctgg gcctgggcga
      901 gccagaggga agcggactgc caccaggacc tgtgctggag gcaagatacg tggcaaggct
      961 gtctgccgca gccgtgctgt atctgagcaa ccctgagggc acctgtgagg atgcaagagc
     1021 aggactgtgg gcatcccacg cagaccacct gctggccctg ctggagtccc caaaggccct
     1081 gacacccggc ctgtcttggc tgctgcagag gatgcaggca agagcagcag gacagacccc
     1141 aaagatggca tgcgtggaca tcccccagct gctggaggag gccgtgggag caggagcacc
     1201 aggaagcgcc ggaggcgtgc tggccgccct gctggatcac gtgcggtctg gcagctgttt
     1261 ccacgccctg ccatcccccc agtactttgt ggacttcgtg tttcagcagc acagctccga
     1321 ggtgccaatg accctggcag agctgagcgc cctgatgcag aggctgggag tgggaaggga
     1381 ggcacactcc gatcactctc acaggcacag gggagcatct agcagggacc ctgtgccact
     1441 gatctcctct agcaattcct ctagcgtgtg ggatacagtg tgcctgtccg ccagagacgt
     1501 gatggcagca tatggactgt ctgagcaggc aggagtgacc cctgaggcat gggcccagct
     1561 gagcccagcc ctgctgcagc agcagctgtc cggagcatgt acatcccagt ctaggcctcc
     1621 agtgcaggat cagctgagcc agtccgagcg gtacctgtat ggctctctgg ccaccctgct
     1681 gatctgcctg tgcgccgtgt tcggactgct gctgctgacc tgcacaggct gtagaggcgt
     1741 gacccactac atcctgcaga catttctgtc cctggcagtg ggagcagtga ccggcgacgc
     1801 cgtgctgcac ctgacaccca aggtgctggg actgcacacc cacagcgagg agggactgtc
     1861 cccacagcct acatggaggc tgctggcaat gctggcagga ctgtatgcct tctttctgtt
     1921 cgagaacctg tttaatctgc tgctgccaag ggaccctgag gatctggagg acggcccttg
     1981 cggacactcc tctcactccc acggaggaca ctcccacggc gtgtctctgc agctggcacc
     2041 ttctgagctg aggcagccaa agccacctca cgagggatct agagccgatc tggtggccga
     2101 ggagagccca gagctgctga acccagagcc acggagactg agcccagagc tgaggctgct
     2161 gccttacatg atcaccctgg gcgatgccgt gcacaatttc gcagacggac tggcagtggg
     2221 agcagccttt gccagctcct ggaagaccgg actggccaca tccctggccg tgttctgcca
     2281 cgagctgcca cacgagctgg gcgactttgc cgccctgctg cacgcaggac tgagcgtgcg
     2341 ccaggccctg ctgctgaacc tggcatccgc cctgaccgcc ttcgccggcc tgtacgtggc
     2401 cctggccgtg ggcgtgagcg aggagtccga ggcatggatc ctggcagtgg caacaggcct
     2461 gttcctgtat gtggccctgt gcgatatgct gcctgccatg ctgaaggtgc gggaccctag
     2521 accatggctg ctgtttctgc tgcacaatgt gggactgctg ggaggatgga cagtgctgct
     2581 gctgctgagc ctgtatgagg acgatatcac ctttgaccca gctttcttgt acaaagttgg
     2641 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2701 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2761 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2821 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2881 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2941 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     3001 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     3061 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     3121 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     3181 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3241 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3301 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3361 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3421 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3481 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3541 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3601 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3661 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3721 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3781 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3841 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3901 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3961 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4021 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4081 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4141 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4201 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4261 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4321 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4381 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4441 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
