LOCUS       sequence-369848-        4998 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 4998)
  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 4998)
  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-SLC9A1_STOP
FEATURES             Location/Qualifiers
     source          1..4998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1318..1332
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     protein_bind    complement(2555..2654)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2671..2690)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2672..2690)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2695..2711)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2695..2711)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2813..2832)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2824..3633
                     /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(2891..2910)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3780..4368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4269..4288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4522..4539
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4698..4725)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4817..4903)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4952..4974
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4966..4983
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4967..4983
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggt gctgcgctct
      121 ggaatctgcg gactgagccc acaccggatc ttcccctccc tgctggtggt ggtggccctg
      181 gtgggactgc tgcctgtgct gagatctcac ggactgcagc tgagcccaac cgcatccaca
      241 atcaggagct ccgagccacc tagggagcgg agcatcggcg acgtgaccac agcaccacca
      301 gaggtgacac ctgagtccag gccagtgaac cactctgtga ccgatcacgg catgaagcct
      361 cggaaggcct tcccagtgct gggcatcgac tacacccacg tgagaacacc ctttgagatc
      421 agcctgtgga tcctgctggc ctgcctgatg aagatcggct tccacgtgat ccccacaatc
      481 tctagcatcg tgcctgagtc ctgtctgctg atcgtggtgg gactgctggt gggaggactg
      541 atcaagggag tgggagagac ccctccattc ctgcagtctg acgtgttctt cctgttcctg
      601 ctgcccccta tcatcctgga cgccggctac ttcctgccac tgaggcagtt taccgagaac
      661 ctgggcacaa tcctgatctt cgccgtggtg ggcacactgt ggaatgcctt ctttctgggc
      721 ggcctgatgt atgccgtgtg cctggtggga ggagagcaga tcaacaatat cggcctgctg
      781 gataacctgc tgtttggcag catcatctcc gccgtggacc cagtggccgt gctggccgtg
      841 ttcgaggaga tccacatcaa cgagctgctg cacatcctgg tgtttggcga gtctctgctg
      901 aatgatgccg tgaccgtggt gctgtaccac ctgttcgagg agtttgccaa ttatgagcac
      961 gtgggcatcg tggacatctt cctgggcttt ctgagcttct ttgtggtggc cctgggaggc
     1021 gtgctggtgg gagtggtgta cggcgtgatc gcagccttca cctctcggtt caccagccac
     1081 atcagagtga tcgagcccct gttcgtgttt ctgtacagct atatggccta tctgtccgcc
     1141 gagctgttcc acctgtctgg catcatggca ctgatcgcaa gcggagtggt catgcggcct
     1201 tacgtggagg ccaatatctc tcacaagagc cacaccacaa tcaagtattt tctgaagatg
     1261 tggtcctctg tgagcgagac actgatcttc atctttctgg gcgtgagcac cgtggcagga
     1321 tcccaccact ggaactggac cttcgtgatc tccacactgc tgttttgcct gatcgcaagg
     1381 gtgctgggcg tgctgggact gacatggttc atcaataagt ttcgcatcgt gaagctgacc
     1441 cccaaggatc agttcatcat cgcatacgga ggactgagag gagcaatcgc cttttccctg
     1501 ggctatctgc tggataagaa gcacttccct atgtgcgacc tgtttctgac cgccatcatc
     1561 acagtgatct tctttacagt gttcgtgcag ggcatgacca tcaggccact ggtggatctg
     1621 ctggccgtga agaagaagca ggagaccaag cgcagcatca acgaggagat ccacacacag
     1681 tttctggatc acctgctgac cggcatcgag gacatctgcg gccactacgg ccaccaccac
     1741 tggaaggaca agctgaaccg gttcaataag aagtatgtga agaagtgtct gatcgccggc
     1801 gagagatcta aggagcctca gctgatcgcc ttttaccaca agatggagat gaagcaggca
     1861 atcgagctgg tggagagcgg aggaatgggc aagatcccat ccgccgtgtc taccgtgagc
     1921 atgcagaata tccaccccaa gtccctgcct tctgagagga tcctgcccgc cctgtccaag
     1981 gataaggagg aggagatccg caagatcctg cggaacaatc tgcagaagac acggcagaga
     2041 ctgaggtctt acaacaggca caccctggtg gccgacccat atgaggaggc ctggaatcag
     2101 atgctgctgc ggagacagaa ggcccgccag ctggagcaga agatcaacaa ttacctgaca
     2161 gtgccagccc acaagctgga ttccccaacc atgtctaggg caaggatcgg atccgaccca
     2221 ctggcatatg agcctaagga ggatctgcca gtgatcacca tcgacccagc aagcccacag
     2281 tcccctgagt ctgtggacct ggtgaacgag gagctgaagg gcaaggtgct gggactgagc
     2341 agggatcctg caaaggtggc agaggaggac gaggacgatg acggaggcat catgatgaga
     2401 tccaaggaga ccagctcccc cggcacagat gacgtgttca ccccagcccc ctccgattct
     2461 ccttctagcc agagaatcca gaggtgcctg agcgaccctg gaccacaccc agagcctgga
     2521 gagggagagc cattctttcc caagggccag taagacccag ctttcttgta caaagttggc
     2581 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     2641 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     2701 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     2761 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     2821 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     2881 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2941 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     3001 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     3061 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     3121 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     3181 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     3241 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     3301 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     3361 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     3421 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     3481 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3541 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3601 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     3661 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     3721 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3781 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3841 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3901 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3961 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4021 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4081 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4141 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4201 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4261 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4321 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4381 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4441 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4501 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4561 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4621 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     4681 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     4741 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     4801 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     4861 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     4921 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     4981 agtcttaagc tcgggccc
//
