LOCUS       sequence-355046-        5316 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 5316)
  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 5316)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..5316
                     /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"
     protein_bind    complement(2873..2972)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2989..3008)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2990..3008)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3013..3029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3013..3029)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3131..3150)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3142..3951
                     /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(3209..3228)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4098..4686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4587..4606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4840..4857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5016..5043)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5135..5221)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5270..5292
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5284..5301
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5285..5301
                     /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 ccaccatggc ccccctggcc
      121 ctggtgggcg tgaccctgct gctggcagca ccaccttgca gcggagcagc aacccctaca
      181 ccatccctgc caccacctcc agcaaacgac agcgatacct ccacaggcgg ctgccagggc
      241 tcttacaggt gtcagccagg cgtgctgctg cccgtgtggg agccagacga tccaagcctg
      301 ggcgacaagg cagcaagggc agtggtgtat ttcgtggcca tggtgtacat gtttctgggc
      361 gtgagcatca tcgccgatcg ctttatggcc gccatcgaag tgatcacatc caaggagaag
      421 gagatcacca tcacaaaggc caatggcgag accagcgtgg gcacagtgcg catctggaac
      481 gagaccgtgt ccaatctgac actgatggca ctgggaagct ccgccccaga gatcctgctg
      541 tccgtgatcg aggtgtgcgg ccacaacttc caggccggcg agctgggacc aggaaccatc
      601 gtgggatccg ccgccttcaa tatgtttgtg gtcatcgccg tgtgcatcta tgtgatcccc
      661 gccggcgagt ctcggaagat caagcacctg agagtgttct ttgtgacagc ctcctggtct
      721 atcttcgcct acgtgtggct gtacctgatc ctggccgtgt ttagccctgg agtggtgcaa
      781 gtgtgggagg ccctgctgac cctggtgttc tttccagtgt gcgtggtgtt cgcctggatg
      841 gccgacaagc ggctgctgtt ttacaagtac gtgtacaagc ggtaccggac agatcctaga
      901 tccggaatca tcatcggagc agagggcgac ccacctaagt ctatcgagct ggatggcacc
      961 tttgtgggag cagaggcacc aggcgagctg ggaggactgg gaccaggacc tgcagaggcc
     1021 cgggagctgg acgcctcccg gagagaagtg atccagatcc tgaaggatct gaagcagaag
     1081 cacccagaca aggatctgga gcagctggtg ggcatcgcca actactatgc cctgctgcac
     1141 cagcagaagt ctagagcctt ctataggatc caggcaacca ggctgatgac aggagcaggc
     1201 aatgtgctga ggcgccacgc cgccgacgcc agcaggagag cagcaccagc agagggagca
     1261 ggcgaggatg aggacgatgg cgcctcccgc atcttctttg agccttctct gtaccactgc
     1321 ctggagaact gtggctctgt gctgctgagc gtgacctgtc agggaggaga gggcaatagc
     1381 accttttatg tggactacag gacagaggat ggctccgcca aggccggctc tgactatgag
     1441 tacagcgagg gcaccctggt gtttaagcct ggcgagacac agaaggagct gcgcatcggc
     1501 atcatcgacg atgacatctt cgaggaggat gagcacttct ttgtgcggct gctgaacctg
     1561 agagtgggcg acgcccaggg catgttcgag ccagatggag gaggcagacc caagggccgg
     1621 ctggtggccc ctctgctggc caccgtgaca atcctggatg acgatcacgc cggcatcttc
     1681 tcttttcagg accggctgct gcacgtgagc gagtgcatgg gcacagtgga tgtgagagtg
     1741 gtgaggtcta gcggagcaag aggaaccgtg aggctgccat atcgcaccgt ggacggaaca
     1801 gcaagaggag gaggagtgca ctacgaggat gcctgtggcg agctggagtt cggcgacgat
     1861 gagaccatga agacactgca ggtgaagatc gtggacgatg aggagtatga gaagaaggac
     1921 aacttcttta tcgagctggg ccagccacag tggctgaaga ggggcatctc cgccctgctg
     1981 ctgaatcagg gcgacggcga taggaagctg accgcagagg aggaggaggc aaggaggatc
     2041 gcagagatgg gcaagcccgt gctgggagag aattgccggc tggaagtgat catcgaggag
     2101 tcttacgact ttaagaacac cgtggataag ctgatcaaga agacaaacct ggccctggtc
     2161 atcggcaccc actcttggag agagcagttc ctggaggcaa tcaccgtgag cgccggcgac
     2221 gaagaggagg aggaggatgg aagcagggag gagcggctgc catcctgctt cgactatgtg
     2281 atgcacttcc tgacagtgtt ttggaaggtg ctgtttgcct gcgtgccacc aaccgagtac
     2341 tgccacggat gggcctgttt cggcgtgtcc atcctggtca tcggcctgct gacagccctg
     2401 atcggcgacc tggcatccca cttcggatgc accgtgggcc tgaaggattc tgtgaacgca
     2461 gtggtgttcg tggccctggg cacctctatc cctgacacat ttgccagcaa ggtggccgcc
     2521 ctgcaggacc agtgtgcaga tgcctccatc ggcaatgtga ccggctctaa cgccgtgaac
     2581 gtgttcctgg gactgggagt ggcatggagc gtggcagccg tgtattgggc agtgcagggc
     2641 aggcccttcg aggtgcgcac cggcacactg gccttctccg tgaccctgtt tacagtgttc
     2701 gcctttgtgg gcatcgccgt gctgctgtac cggagaaggc ctcacatcgg aggagagctg
     2761 ggaggaccta ggggaccaaa gctggcaacc acagccctgt tcctgggcct gtggctgctg
     2821 tatatcctgt ttgccagcct ggaggcctac tgtcacatca gaggcttcta agacccagct
     2881 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     2941 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     3001 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     3061 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     3121 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     3181 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     3241 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     3301 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     3361 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     3421 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     3481 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     3541 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     3601 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3661 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     3721 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3781 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3841 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3901 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     3961 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     4021 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     4081 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4681 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4741 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4801 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4861 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4921 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     4981 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     5041 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     5101 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     5161 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     5221 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     5281 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
