LOCUS       sequence-259222-        5292 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 5292)
  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 5292)
  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_SLC12A9
FEATURES             Location/Qualifiers
     source          1..5292
                     /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(3417..3516)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3533..3552)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3534..3552)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3557..3573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3557..3573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3675..3694)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3686..4495
                     /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(3753..3772)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4642..5230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5131..5150
                     /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 accatggcca gcgagagctc ccctctgctg gcatacaggc tgctgggaga
      721 ggagggcgtg gccctgcctg ccaatggagc aggcggacca ggaggagcat ctgccagaaa
      781 gctgagcacc tttctgggcg tggtggtgcc aacagtgctg tctatgttca gcatcgtggt
      841 gtttctgcgg atcggcttcg tggtgggaca cgcaggactg ctgcaggccc tggcaatgct
      901 gctggtggcc tatttcatcc tggccctgac cgtgctgtcc gtgtgcgcca tcgcaacaaa
      961 tggagcagtg cagggaggag gcgcctactt tatgatcagc agaaccctgg gacctgaagt
     1021 gggaggatcc atcggcctga tgttctatct ggccaacgtg tgcggatgtg ccgtgagcct
     1081 gctgggactg gtggagtccg tgctggacgt gtttggagca gatgcaacag gaccatccgg
     1141 actgagggtg ctgcctcagg gatacggctg gaatctgctg tatggctctc tgctgctggg
     1201 cctggtggga ggcgtgtgca ccctgggagc aggcctgtac gcccgcgcca gcttcctgac
     1261 atttctgctg gtgtctggca gcctggccag cgtgctgatc tccttcgtgg cagtgggacc
     1321 acgggatatc agactgaccc ctaggccagg accaaacgga tctagcctgc cccctagatt
     1381 cggccacttt accggcttca actcctctac actgaaggac aatctgggcg ccggctacgc
     1441 cgaggattat accacaggcg ccgtgatgaa ctttgcctcc gtgttcgccg tgctgtttaa
     1501 tggctgcaca ggcatcatgg ccggcgccaa catgtctggc gagctgaagg accctagcag
     1561 ggcaatccca ctgggaacca tcgtggcagt ggcctacacc ttcttcgtgt acgtgctgct
     1621 gttctttctg agctccttta cctgtgacag gacactgctg caggaggatt acggcttctt
     1681 tcgcgccatc tctctgtggc cacccctggt gctgatcggc atctatgcca ccgccctgtc
     1741 cgcctctatg tctagcctga tcggagcaag caggatcctg cacgcactgg caagagacga
     1801 tctgttcggc gtgatcctgg cacctgcaaa ggtggtgtcc aggggaggaa atccatgggc
     1861 agccgtgctg tactcttggg gactggtgca gctggtgctg ctggcaggca agctgaacac
     1921 cctggcagcc gtggtgacag tgttttacct ggtggcctat gccgccgtgg atctgtcctg
     1981 cctgtctctg gagtgggcaa gcgccccaaa tttcaggccc acctttagcc tgttctcctg
     2041 gcacacatgc ctgctgggcg tggcctcctg tctgctgatg atgttcctga tctccccagg
     2101 agcagcagga ggatctctgc tgctgatggg actgctggcc gccctgctga ccgcaagggg
     2161 aggaccttcc tcttggggct acgtgagcca ggccctgctg tttcaccagg tgcggaagta
     2221 tctgctgagg ctggacgtgc gcaaggatca cgtgaagttc tggagacccc agctgctgct
     2281 gctggtgggc aatccaaggg gcgccctgcc tctgctgagg ctggccaacc agctgaagaa
     2341 gggaggactg tacgtgctgg gacacgtgac cctgggcgac ctggattctc tgcctagcga
     2401 tccagtgcag ccccagtatg gagcatggct gagcctggtg gacagggcac aggtgaaggc
     2461 ctttgtggat ctgacactga gcccatccgt gcggcaggga gcacagcacc tgctgagaat
     2521 ctccggactg ggaggaatga agccaaacac cctggtgctg ggcttctacg acgatgcccc
     2581 tccacaggac cactttctga cagatccagc cttctccgag ccagcagact ctaccaggga
     2641 gggcagctcc cctgccctgt ctacactgtt cccacctcca agggcacctg gaagcccacg
     2701 cgccctgaat ccacaggact atgtggccac cgtggccgat gccctgaaga tgaacaagaa
     2761 tgtggtgctg gcaagagcat ctggcgccct gccacctgag aggctgtcca ggggatctgg
     2821 aggcacaagc cagctgcacc acgtggacgt gtggccactg aacctgctga ggcctagagg
     2881 aggaccagga tacgtggacg tgtgcggact gtttctgctg cagatggcaa ccatcctggg
     2941 aatggtgcca gcatggcact ccgccaggct gcgcatcttc ctgtgcctgg gaccaagaga
     3001 ggcacctgga gcagcagagg gccggctgag agccctgctg tctcagctga ggatccgcgc
     3061 cgaggtgcag gaggtggtgt ggggagaggg agcaggagca ggagagccag aggccgagga
     3121 ggagggcgac tttgtgaata gcggcagggg cgatgccgag gccgaggccc tggcccgctc
     3181 cgccaacgcc ctggtgcggg cccagcaggg cagaggcacc ggcggaggac caggaggacc
     3241 tgagggaggc gacgccgagg gccccatcac cgccctgaca ttcctgtatc tgcctaggcc
     3301 accagcagac ccagcaaggt accccagata tctggccctg ctggagaccc tgacacgcga
     3361 tctgggacca accctgctgg tgcacggagt gacacccgtg acctgtacag acctggaccc
     3421 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     3481 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     3541 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     3601 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     3661 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     3721 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     3781 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     3841 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3901 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3961 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     4021 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     4081 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     4141 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     4201 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     4261 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     4321 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     4381 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     4441 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     4501 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     4561 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     4621 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4681 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4741 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4801 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4861 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4921 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4981 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5041 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5101 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5161 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5221 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5281 tgctcacatg tt
//
