LOCUS       sequence-438927-        4296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4296)
  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 4296)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4296
                     /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(2421..2520)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2537..2556)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2538..2556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2561..2577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2561..2577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2679..2698)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2690..3499
                     /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(2757..2776)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3646..4234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4135..4154
                     /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 ctgctgtaag acccagcttt cttgtacaaa gttggcatta taagaaagca
     2461 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2521 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2581 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2641 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2701 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2761 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2821 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2881 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2941 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3001 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3061 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3121 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3181 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3241 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3301 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3361 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3421 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3481 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3541 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3781 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4261 gttcctggcc ttttgctggc cttttgctca catgtt
//
