LOCUS       sequence-263916-        4944 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 4944)
  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 4944)
  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_SLC9A4
FEATURES             Location/Qualifiers
     source          1..4944
                     /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(3069..3168)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3185..3204)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3186..3204)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3209..3225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3209..3225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3327..3346)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3338..4147
                     /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(3405..3424)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4294..4882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4783..4802
                     /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 accatggccc tgcagatgtt cgtgacctac tctccctgga actgcctgct
      721 gctgctggtg gccctggagt gtagcgaggc cagctccgat ctgaacgagt ctgccaatag
      781 cacagcccag tatgccagca atgcatggtt cgcagcagca tctagcgagc cagaggaggg
      841 catctccgtg tttgagctgg attacgacta tgtgcagatc ccctacgagg tgaccctgtg
      901 gatcctgctg gcctctctgg ccaagatcgg cttccacctg tatcacaggc tgcctggcct
      961 gatgccagag agctgcctgc tgatcctggt gggcgccctg gtgggaggca tcatctttgg
     1021 caccgatcac aagtcccccc ctgtgatgga ctcctctatc tacttcctgt atctgctgcc
     1081 accaatcgtg ctggagggag gatactttat gcccacacgc cctttctttg agaacatcgg
     1141 ctccatcctg tggtgggccg tgctgggcgc cctgatcaat gcactgggca tcggcctgtc
     1201 tctgtatctg atctgtcagg tgaaggcctt cggcctgggc gatgtgaacc tgctgcagaa
     1261 tctgctgttt ggctccctga tctctgccgt ggaccctgtg gccgtgctgg ccgtgttcga
     1321 ggaggcacgg gtgaacgagc agctgtacat gatgatcttt ggcgaggccc tgctgaacga
     1381 cggcatcacc gtggtgctgt ataatatgct gatcgccttc accaagatgc acaagtttga
     1441 ggatatcgag acagtggaca tcctggcagg atgcgcaagg ttcatcgtgg tgggactggg
     1501 aggcgtgctg tttggcatcg tgttcggctt tatctccgcc ttcatcaccc gctttacaca
     1561 gaacatctct gccatcgagc ccctgatcgt gttcatgttt agctacctgt cctatctggc
     1621 agcagagacc ctgtacctgt ccggaatcct ggcaatcacc gcatgtgcag tgacaatgaa
     1681 gaagtatgtg gaggagaacg tgagccagac aagctacacc acaatcaagt atttcatgaa
     1741 gatgctgagc tccgtgagcg agaccctgat cttcatcttt atgggcgtgt ccacagtggg
     1801 caagaaccac gagtggaatt gggccttcat ctgctttacc ctggcctttt gtcagatctg
     1861 gcgggccatc agcgtgttcg ccctgtttta catcagcaac cagttcagaa cattcccttt
     1921 ttctatcaag gatcagtgca tcatctttta tagcggcgtg aggggagcag gatctttcag
     1981 cctggccttt ctgctgcctc tgagcctgtt cccacgcaag aagatgtttg tgaccgccac
     2041 actggtggtc atctacttca ccgtgttcat ccagggcatc acagtgggcc cactggtgcg
     2101 gtatctggac gtgaagaaga ccaacaagaa ggagtccatc aatgaggagc tgcacatcag
     2161 actgatggat cacctgaagg caggaatcga ggacgtgtgc ggacactgga gccactacca
     2221 ggtgagggat aagttcaaga agtttgacca caggtatctg cgcaagatcc tgatccgcaa
     2281 gaatctgccc aagtctagca tcgtgtccct gtacaagaag ctggagatga agcaggccat
     2341 cgagatggtg gagaccggca tcctgtcctc tacagccttc agcatccccc accaggccca
     2401 gaggatccag ggcatcaagc gcctgtctcc tgaggatgtg gagagcatca gagacatcct
     2461 gacctctaac atgtaccagg tgcggcagag aacactgagc tacaacaagt ataatctgaa
     2521 gcctcagacc tccgagaagc aggccaagga gatcctgatc cggagacaga atacactgag
     2581 ggagtccatg cgcaagggac actctctgcc atggggcaag cctgcaggaa ccaagaacat
     2641 ccggtacctg tcctacccat atggcaatcc ccagtctgcc ggaagggata ccagagcagc
     2701 aggcttcagc gacgatgaca gctccgaccc aggctccccc tctatcacat tttccgcctg
     2761 ctctagaatc ggctccctgc agaagcagga ggcccaggag atcatcccaa tgaagtctct
     2821 gcacaggggc cgcaaggcct tcagctttgg ctaccagcgg aacacctccc aggaggagta
     2881 tctgggaggc gtgaggaggg tggccctgag acctaagcca ctgttccacg cagtggatga
     2941 ggagggagag agcggaggag agtccgaggg caaggcaagc ctggtggagg tgcggtccag
     3001 atggacagca gatcacggac acggaaggga ccaccacaga agccactccc ccctgctgca
     3061 gaagaaggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     3121 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     3181 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3241 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     3301 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     3361 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3421 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     3481 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3541 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3601 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3661 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3721 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3781 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3841 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3901 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3961 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     4021 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     4081 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     4141 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     4201 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     4261 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4321 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4381 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4441 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4501 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4561 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4621 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4681 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4741 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4801 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4861 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4921 ttgctggcct tttgctcaca tgtt
//
