LOCUS       sequence-261171-        4293 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 4293)
  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 4293)
  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_SLC9A8
FEATURES             Location/Qualifiers
     source          1..4293
                     /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(2418..2517)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2534..2553)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2535..2553)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2558..2574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2558..2574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2676..2695)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2687..3496
                     /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(2754..2773)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3643..4231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4132..4151
                     /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 accatgggcg agaagatggc cgaggaggag cggttcccaa acaccacaca
      721 cgagggcttt aatgtgacac tgcacaccac actggtggtg accacaaagc tggtgctgcc
      781 aacacccggc aagcctatcc tgccagtgca gaccggagag caggcacagc aggaggagca
      841 gagctccggc atgaccatct tctttagcct gctggtgctg gccatctgca tcatcctggt
      901 gcacctgctg atcaggtaca ggctgcactt cctgccagag tccgtggcag tggtgtctct
      961 gggcatcctg atgggcgccg tgatcaagat catcgagttc aagaagcttg ccaactggaa
     1021 agaggaggag atgtttagac ctaacatgtt ctttctgctg ctgctgcccc ctatcatctt
     1081 cgagagcggc tactccctgc acaagggcaa cttctttcag aatatcggct ccatcacact
     1141 gttcgccgtg tttggcaccg caatctctgc cttcgtggtg ggaggaggca tctattttct
     1201 gggccaggcc gacgtgatct ctaagctgaa tatgacagat agcttcgcct ttggctctct
     1261 gatcagcgcc gtggacccag tggcaaccat cgccatcttc aacgccctgc acgtggatcc
     1321 cgtgctgaat atgctggtgt ttggcgagtc catcctgaac gacgccgtgt ctatcgtgct
     1381 gacaaatacc gccgagggcc tgacaaggaa gaacatgtcc gacgtgtctg gctggcagac
     1441 cttcctgcag gccctggatt actttctgaa gatgttcttt ggaagcgccg ccctgggcac
     1501 actgaccgga ctgatctccg ccctggtgct gaagcacatc gatctgcgca agacccctag
     1561 cctggagttc ggcatgatga tcatctttgc ctacctgcca tatggactgg cagagggaat
     1621 ctctctgagc ggaatcatgg ccatcctgtt cagcggcatc gtgatgtccc actatacaca
     1681 ccacaacctg tccccagtga cccagatcct gatgcagcag acactgagga ccgtggcctt
     1741 tctgtgcgag acctgcgtgt tcgcctttct gggcctgagc atcttctcct ttccccacaa
     1801 gttcgagatc agctttgtga tctggtgtat cgtgctggtg ctgttcggcc gggccgtgaa
     1861 tatctttccc ctgtcctacc tgctgaactt ctttagagac cacaagatca cccctaagat
     1921 gatgttcatc atgtggtttt ctggactgag gggagcaatc ccatatgcac tgagcctgca
     1981 cctggatctg gagcctatgg agaagagaca gctgatcggc accacaacca tcgtgatcgt
     2041 gctgttcaca atcctgctgc tgggcggctc taccatgcct ctgatcagac tgatggacat
     2101 cgaggatgcc aaggcccacc ggagaaacaa gaaggacgtg aatctgtcca agacagagaa
     2161 gatgggcaat accgtggagt ctgagcacct gagcgagctg accgaggagg agtacgaggc
     2221 ccactatatc aggcgccagg acctgaaggg cttcgtgtgg ctggatgcca agtacctgaa
     2281 tcctttcttt acacggagac tgacccagga ggatctgcac cacggcagga tccagatgaa
     2341 gacactgacc aacaagtggt atgaggaggt gcgccaggga ccatccggat ctgaggacga
     2401 tgagcaggag ctgctggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2461 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2521 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2581 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2641 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2701 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2761 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2821 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2881 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2941 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     3001 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3061 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3121 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3181 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3241 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3301 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3361 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3421 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3481 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3541 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3601 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3661 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3721 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3781 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3841 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3901 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3961 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4021 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4081 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4141 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4201 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4261 cctggccttt tgctggcctt ttgctcacat gtt
//
