LOCUS       sequence-261071-        5313 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 5313)
  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 5313)
  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_SLC8A2
FEATURES             Location/Qualifiers
     source          1..5313
                     /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(3438..3537)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3554..3573)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3555..3573)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3578..3594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3578..3594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3696..3715)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3707..4516
                     /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(3774..3793)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4663..5251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5152..5171
                     /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 ccctggccct ggtgggcgtg accctgctgc tggcagcacc
      721 accttgcagc ggagcagcaa cccctacacc atccctgcca ccacctccag caaacgacag
      781 cgatacctcc acaggcggct gccagggctc ttacaggtgt cagccaggcg tgctgctgcc
      841 cgtgtgggag ccagacgatc caagcctggg cgacaaggca gcaagggcag tggtgtattt
      901 cgtggccatg gtgtacatgt ttctgggcgt gagcatcatc gccgatcgct ttatggccgc
      961 catcgaagtg atcacatcca aggagaagga gatcaccatc acaaaggcca atggcgagac
     1021 cagcgtgggc acagtgcgca tctggaacga gaccgtgtcc aatctgacac tgatggcact
     1081 gggaagctcc gccccagaga tcctgctgtc cgtgatcgag gtgtgcggcc acaacttcca
     1141 ggccggcgag ctgggaccag gaaccatcgt gggatccgcc gccttcaata tgtttgtggt
     1201 catcgccgtg tgcatctatg tgatccccgc cggcgagtct cggaagatca agcacctgag
     1261 agtgttcttt gtgacagcct cctggtctat cttcgcctac gtgtggctgt acctgatcct
     1321 ggccgtgttt agccctggag tggtgcaagt gtgggaggcc ctgctgaccc tggtgttctt
     1381 tccagtgtgc gtggtgttcg cctggatggc cgacaagcgg ctgctgtttt acaagtacgt
     1441 gtacaagcgg taccggacag atcctagatc cggaatcatc atcggagcag agggcgaccc
     1501 acctaagtct atcgagctgg atggcacctt tgtgggagca gaggcaccag gcgagctggg
     1561 aggactggga ccaggacctg cagaggcccg ggagctggac gcctcccgga gagaagtgat
     1621 ccagatcctg aaggatctga agcagaagca cccagacaag gatctggagc agctggtggg
     1681 catcgccaac tactatgccc tgctgcacca gcagaagtct agagccttct ataggatcca
     1741 ggcaaccagg ctgatgacag gagcaggcaa tgtgctgagg cgccacgccg ccgacgccag
     1801 caggagagca gcaccagcag agggagcagg cgaggatgag gacgatggcg cctcccgcat
     1861 cttctttgag ccttctctgt accactgcct ggagaactgt ggctctgtgc tgctgagcgt
     1921 gacctgtcag ggaggagagg gcaatagcac cttttatgtg gactacagga cagaggatgg
     1981 ctccgccaag gccggctctg actatgagta cagcgagggc accctggtgt ttaagcctgg
     2041 cgagacacag aaggagctgc gcatcggcat catcgacgat gacatcttcg aggaggatga
     2101 gcacttcttt gtgcggctgc tgaacctgag agtgggcgac gcccagggca tgttcgagcc
     2161 agatggagga ggcagaccca agggccggct ggtggcccct ctgctggcca ccgtgacaat
     2221 cctggatgac gatcacgccg gcatcttctc ttttcaggac cggctgctgc acgtgagcga
     2281 gtgcatgggc acagtggatg tgagagtggt gaggtctagc ggagcaagag gaaccgtgag
     2341 gctgccatat cgcaccgtgg acggaacagc aagaggagga ggagtgcact acgaggatgc
     2401 ctgtggcgag ctggagttcg gcgacgatga gaccatgaag acactgcagg tgaagatcgt
     2461 ggacgatgag gagtatgaga agaaggacaa cttctttatc gagctgggcc agccacagtg
     2521 gctgaagagg ggcatctccg ccctgctgct gaatcagggc gacggcgata ggaagctgac
     2581 cgcagaggag gaggaggcaa ggaggatcgc agagatgggc aagcccgtgc tgggagagaa
     2641 ttgccggctg gaagtgatca tcgaggagtc ttacgacttt aagaacaccg tggataagct
     2701 gatcaagaag acaaacctgg ccctggtcat cggcacccac tcttggagag agcagttcct
     2761 ggaggcaatc accgtgagcg ccggcgacga agaggaggag gaggatggaa gcagggagga
     2821 gcggctgcca tcctgcttcg actatgtgat gcacttcctg acagtgtttt ggaaggtgct
     2881 gtttgcctgc gtgccaccaa ccgagtactg ccacggatgg gcctgtttcg gcgtgtccat
     2941 cctggtcatc ggcctgctga cagccctgat cggcgacctg gcatcccact tcggatgcac
     3001 cgtgggcctg aaggattctg tgaacgcagt ggtgttcgtg gccctgggca cctctatccc
     3061 tgacacattt gccagcaagg tggccgccct gcaggaccag tgtgcagatg cctccatcgg
     3121 caatgtgacc ggctctaacg ccgtgaacgt gttcctggga ctgggagtgg catggagcgt
     3181 ggcagccgtg tattgggcag tgcagggcag gcccttcgag gtgcgcaccg gcacactggc
     3241 cttctccgtg accctgttta cagtgttcgc ctttgtgggc atcgccgtgc tgctgtaccg
     3301 gagaaggcct cacatcggag gagagctggg aggacctagg ggaccaaagc tggcaaccac
     3361 agccctgttc ctgggcctgt ggctgctgta tatcctgttt gccagcctgg aggcctactg
     3421 tcacatcaga ggcttcgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     3481 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     3541 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     3601 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     3661 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     3721 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     3781 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     3841 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     3901 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     3961 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     4021 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     4081 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     4141 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     4201 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     4261 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     4321 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     4381 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     4441 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     4501 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     4561 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     4621 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4681 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4741 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4801 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4861 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4921 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4981 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5041 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5101 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5161 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5221 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5281 cctggccttt tgctggcctt ttgctcacat gtt
//
