LOCUS       sequence-261106-        5331 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 5331)
  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 5331)
  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_SLC8A3
FEATURES             Location/Qualifiers
     source          1..5331
                     /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(3456..3555)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3572..3591)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3573..3591)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3596..3612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3596..3612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3714..3733)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3725..4534
                     /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(3792..3811)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4681..5269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5170..5189
                     /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 accatggcct ggctgagact gcagcccctg acaagcgcct ttctgcactt
      721 cggcctggtg acctttgtgc tgttcctgaa cggactgagg gcagaggcag gaggatccgg
      781 cgacgtgcct tctaccggcc agaacaatga gagctgttcc ggcagctccg attgcaagga
      841 gggcgtgatc ctgccaatct ggtaccccga gaatccttcc ctgggcgaca agatcgcccg
      901 cgtgatcgtg tactttgtgg ccctgatcta tatgttcctg ggcgtgtcta tcatcgccga
      961 tcgcttcatg gccagcatcg aagtgatcac atcccaggag cgggaggtga ccatcaagaa
     1021 gccaaacggc gagacatcca ccacaaccat ccgggtgtgg aacgagacag tgtctaatct
     1081 gaccctgatg gcactgggat ctagcgcccc agagatcctg ctgtccctga tcgaggtgtg
     1141 cggacacggc ttcatcgcag gcgacctggg acctagcacc atcgtgggat ccgccgcctt
     1201 taatatgttc atcatcatcg gcatctgcgt gtacgtgatc cccgatggcg agacaagaaa
     1261 gatcaagcac ctgagggtgt tctttatcac cgccgcctgg tccatctttg cctacatctg
     1321 gctgtatatg atcctggccg tgttctctcc tggagtggtg caagtgtggg agggactgct
     1381 gaccctgttc tttttccccg tgtgcgtgct gctggcatgg gtggcagaca agcgcctgct
     1441 gttctacaag tatatgcaca agaagtatcg cacagataag caccggggca tcatcatcga
     1501 gaccgagggc gatcacccaa agggcatcga gatggacggc aagatgatga acagccactt
     1561 tctggatggc aatctggtgc ccctggaggg caaggaggtg gacgagtccc ggagagagat
     1621 gatcagaatc ctgaaggatc tgaagcagaa gcaccccgag aaggacctgg atcagctggt
     1681 ggagatggcc aactactatg ccctgtctca ccagcagaag agcagagcct tctacaggat
     1741 ccaggccaca aggatgatga ccggcgccgg caatatcctg aagaagcacg cagcagagca
     1801 ggccaagaag gcatcctcta tgagcgaggt gcacacagat gagcccgagg actttatctc
     1861 caaggtgttt ttcgaccctt gctcttacca gtgtctggag aactgcggag ccgtgctgct
     1921 gaccgtggtg cggaagggcg gcgacatgtc taagacaatg tacgtggatt ataagaccga
     1981 ggacggaagc gccaatgcag gagcagatta tgagttcaca gagggaaccg tggtgctgaa
     2041 gccaggagag acccagaagg agttttctgt gggcatcatc gacgatgaca tcttcgagga
     2101 ggacgagcac tttttcgtga gactgagcaa cgtgaggatc gaggaggagc agccagagga
     2161 gggaatgcca cctgccatct ttaatagcct gcctctgcca agagccgtgc tggcatcccc
     2221 ttgcgtggca acagtgacca tcctggatga cgatcacgcc ggcatcttta cattcgagtg
     2281 cgataccatc cacgtgtctg agagcatcgg cgtgatggag gtgaaggtgc tgcgcacaag
     2341 cggagcaagg ggaaccgtga tcgtgccttt caggacagtg gagggaaccg caaagggagg
     2401 aggagaggat tttgaggaca cctacggcga gctggagttc aagaacgacg agacagtgaa
     2461 gaccatccgc gtgaagatcg tggatgagga ggagtatgag cggcaggaga acttcttcat
     2521 cgccctgggc gagccaaagt ggatggagag aggcatctcc gccctgctgc tgtctcccga
     2581 tgtgacagac agaaagctga ccatggagga ggaggaggcc aagaggatcg ccgagatggg
     2641 caagccagtg ctgggcgagc accccaagct ggaagtgatc atcgaggagt cctacgagtt
     2701 caagacaacc gtggacaagc tgatcaagaa gacaaacctg gccctggtgg tgggaaccca
     2761 cagctggcgc gatcagttta tggaggcaat cacagtgtcc gccgcaggcg acgaggatga
     2821 ggacgagtct ggcgaggagc ggctgcctag ctgttttgac tacgtgatgc actttctgac
     2881 agtgttctgg aaggtgctgt tcgcctgcgt gccacccacc gagtattgtc acggctgggc
     2941 ctgctttgcc gtgtctatcc tgatcatcgg catgctgacc gccatcatcg gcgacctggc
     3001 ctctcacttc ggctgtacaa tcggcctgaa ggatagcgtg accgccgtgg tgtttgtggc
     3061 cttcggcaca tccgtgccag acacctttgc ctctaaggca gccgccctgc aggacgtgta
     3121 cgcagacgcc agcatcggca atgtgacagg ctccaacgcc gtgaacgtgt tcctgggaat
     3181 cggactggca tggtccgtgg cagccatcta ttgggccctg cagggccagg agtttcacgt
     3241 gtctgccggc accctggcct ttagcgtgac actgttcacc atctttgcct tcgtgtgcat
     3301 cagcgtgctg ctgtacaggc gccggcctca cctgggagga gagctgggag gaccaagggg
     3361 ctgcaagctg gccacaacct ggctgttcgt gagcctgtgg ctgctgtata tcctgttcgc
     3421 caccctggag gcctactgct atatcaaggg cttcgaccca gctttcttgt acaaagttgg
     3481 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     3541 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     3601 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     3661 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     3721 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     3781 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     3841 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     3901 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     3961 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     4021 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     4081 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     4141 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     4201 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     4261 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     4321 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     4381 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     4441 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     4501 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     4561 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     4621 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4681 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4741 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4801 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4861 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4921 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4981 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5041 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5101 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5161 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5221 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5281 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
