LOCUS       sequence-261211-        4995 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 4995)
  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 4995)
  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
FEATURES             Location/Qualifiers
     source          1..4995
                     /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"
     CDS             1886..1900
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     protein_bind    complement(3120..3219)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3236..3255)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3237..3255)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3260..3276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3260..3276)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3378..3397)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3389..4198
                     /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(3456..3475)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4345..4933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4834..4853
                     /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 accatggtgc tgcgctctgg aatctgcgga ctgagcccac accggatctt
      721 cccctccctg ctggtggtgg tggccctggt gggactgctg cctgtgctga gatctcacgg
      781 actgcagctg agcccaaccg catccacaat caggagctcc gagccaccta gggagcggag
      841 catcggcgac gtgaccacag caccaccaga ggtgacacct gagtccaggc cagtgaacca
      901 ctctgtgacc gatcacggca tgaagcctcg gaaggccttc ccagtgctgg gcatcgacta
      961 cacccacgtg agaacaccct ttgagatcag cctgtggatc ctgctggcct gcctgatgaa
     1021 gatcggcttc cacgtgatcc ccacaatctc tagcatcgtg cctgagtcct gtctgctgat
     1081 cgtggtggga ctgctggtgg gaggactgat caagggagtg ggagagaccc ctccattcct
     1141 gcagtctgac gtgttcttcc tgttcctgct gccccctatc atcctggacg ccggctactt
     1201 cctgccactg aggcagttta ccgagaacct gggcacaatc ctgatcttcg ccgtggtggg
     1261 cacactgtgg aatgccttct ttctgggcgg cctgatgtat gccgtgtgcc tggtgggagg
     1321 agagcagatc aacaatatcg gcctgctgga taacctgctg tttggcagca tcatctccgc
     1381 cgtggaccca gtggccgtgc tggccgtgtt cgaggagatc cacatcaacg agctgctgca
     1441 catcctggtg tttggcgagt ctctgctgaa tgatgccgtg accgtggtgc tgtaccacct
     1501 gttcgaggag tttgccaatt atgagcacgt gggcatcgtg gacatcttcc tgggctttct
     1561 gagcttcttt gtggtggccc tgggaggcgt gctggtggga gtggtgtacg gcgtgatcgc
     1621 agccttcacc tctcggttca ccagccacat cagagtgatc gagcccctgt tcgtgtttct
     1681 gtacagctat atggcctatc tgtccgccga gctgttccac ctgtctggca tcatggcact
     1741 gatcgcaagc ggagtggtca tgcggcctta cgtggaggcc aatatctctc acaagagcca
     1801 caccacaatc aagtattttc tgaagatgtg gtcctctgtg agcgagacac tgatcttcat
     1861 ctttctgggc gtgagcaccg tggcaggatc ccaccactgg aactggacct tcgtgatctc
     1921 cacactgctg ttttgcctga tcgcaagggt gctgggcgtg ctgggactga catggttcat
     1981 caataagttt cgcatcgtga agctgacccc caaggatcag ttcatcatcg catacggagg
     2041 actgagagga gcaatcgcct tttccctggg ctatctgctg gataagaagc acttccctat
     2101 gtgcgacctg tttctgaccg ccatcatcac agtgatcttc tttacagtgt tcgtgcaggg
     2161 catgaccatc aggccactgg tggatctgct ggccgtgaag aagaagcagg agaccaagcg
     2221 cagcatcaac gaggagatcc acacacagtt tctggatcac ctgctgaccg gcatcgagga
     2281 catctgcggc cactacggcc accaccactg gaaggacaag ctgaaccggt tcaataagaa
     2341 gtatgtgaag aagtgtctga tcgccggcga gagatctaag gagcctcagc tgatcgcctt
     2401 ttaccacaag atggagatga agcaggcaat cgagctggtg gagagcggag gaatgggcaa
     2461 gatcccatcc gccgtgtcta ccgtgagcat gcagaatatc caccccaagt ccctgccttc
     2521 tgagaggatc ctgcccgccc tgtccaagga taaggaggag gagatccgca agatcctgcg
     2581 gaacaatctg cagaagacac ggcagagact gaggtcttac aacaggcaca ccctggtggc
     2641 cgacccatat gaggaggcct ggaatcagat gctgctgcgg agacagaagg cccgccagct
     2701 ggagcagaag atcaacaatt acctgacagt gccagcccac aagctggatt ccccaaccat
     2761 gtctagggca aggatcggat ccgacccact ggcatatgag cctaaggagg atctgccagt
     2821 gatcaccatc gacccagcaa gcccacagtc ccctgagtct gtggacctgg tgaacgagga
     2881 gctgaagggc aaggtgctgg gactgagcag ggatcctgca aaggtggcag aggaggacga
     2941 ggacgatgac ggaggcatca tgatgagatc caaggagacc agctcccccg gcacagatga
     3001 cgtgttcacc ccagccccct ccgattctcc ttctagccag agaatccaga ggtgcctgag
     3061 cgaccctgga ccacacccag agcctggaga gggagagcca ttctttccca agggccagga
     3121 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     3181 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     3241 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     3301 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     3361 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     3421 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     3481 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     3541 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     3601 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     3661 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     3721 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3781 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3841 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3901 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3961 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     4021 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     4081 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     4141 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     4201 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     4261 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     4321 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4381 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4441 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4501 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4561 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4621 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4681 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4741 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4801 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4861 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4921 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4981 ttttgctcac atgtt
//
