LOCUS       sequence-262891-        4161 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 4161)
  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 4161)
  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_SLC9B2
FEATURES             Location/Qualifiers
     source          1..4161
                     /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(2286..2385)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2402..2421)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2403..2421)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2426..2442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2426..2442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2544..2563)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2555..3364
                     /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(2622..2641)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3511..4099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4000..4019
                     /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 acgaggataa gcggatcacc tacgaggact ccgagccatc
      721 taccggcatg aactatacac cctctatgca ccaggaggcc caggaggaga ccgtgatgaa
      781 gctgaagggc atcgatgcca atgagccaac agagggcagc atcctgctga agagctccga
      841 gaagaagctg caggagaccc caacagaggc aaaccacgtg cagcggctga gacagatgct
      901 ggcatgccca cctcacggac tgctggacag agtgatcacc aatgtgacaa tcatcgtgct
      961 gctgtgggcc gtggtgtggt ccatcaccgg atctgagtgc ctgccaggag gaaacctgtt
     1021 cggcatcatc atcctgtttt actgtgccat catcggcggc aagctgctgg gcctgatcaa
     1081 gctgcccaca ctgccacccc tgccttccct gctgggaatg ctgctggcag gcttcctgat
     1141 caggaatatc cccgtgatca acgacaatgt gcagatcaag cacaagtggt ctagctccct
     1201 gcggagcatc gccctgtcca tcatcctggt gagagccggc ctgggcctgg attctaaggc
     1261 cctgaagaag ctgaagggcg tgtgcgtgcg cctgtctatg ggaccttgca tcgtggaggc
     1321 atgtaccagc gccctgctgg cacactacct gctgggactg ccatggcagt ggggcttcat
     1381 cctgggcttt gtgctgggag ccgtgtctcc tgcagtggtg gtgccaagca tgctgctgct
     1441 gcagggagga ggatatggag tggagaaggg agtgcctaca ctgctgatgg cagcaggctc
     1501 cttcgacgat atcctggcca tcaccggctt taacacatgt ctgggcatcg ccttcagcac
     1561 cggctccaca gtgtttaatg tgctgagggg cgtgctggag gtggtcatcg gagtggcaac
     1621 cggatccgtg ctgggcttct ttatccagta cttcccctct agggaccagg ataagctggt
     1681 gtgcaagcgc acattcctgg tgctgggact gagcgtgctg gccgtgtttt ctagcgtgca
     1741 cttcggattt cctggatccg gaggactgtg caccctggtc atggcctttc tggccggcat
     1801 gggctggaca tccgagaagg ccgaggtgga gaagatcatc gccgtggcct gggatatctt
     1861 ccagcccctg ctgtttggcc tgatcggcgc cgaggtgtct atcgcaagcc tgaggcctga
     1921 gaccgtggga ctgtgcgtgg caacagtggg aatcgccgtg ctgatccgca tcctgaccac
     1981 attcctgatg gtgtgcttcg ccggctttaa cctgaaggag aagatcttca tcagctttgc
     2041 ctggctgccc aaggccaccg tgcaggcagc aatcggatcc gtggccctgg acacagcacg
     2101 gtctcacggc gagaagcagc tggaggacta tggcatggat gtgctgaccg tggcctttct
     2161 gagcatcctg atcacagccc ctatcggatc cctgctgatc ggactgctgg gaccaagact
     2221 gctgcagaag gtggagcacc agaataagga tgaggaggtg cagggcgaga ccagcgtgca
     2281 ggtggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2341 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2401 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2461 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2521 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2581 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2641 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2701 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2761 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2821 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2881 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2941 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3001 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3061 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3121 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3181 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3241 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3301 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3361 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3421 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4141 ctggcctttt gctcacatgt t
//
