LOCUS       sequence-353418-        4179 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 4179)
  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 4179)
  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-SLC22A8_STOP
FEATURES             Location/Qualifiers
     source          1..4179
                     /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(1736..1835)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1852..1871)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1853..1871)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1876..1892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1876..1892)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1994..2013)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2005..2814
                     /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(2072..2091)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2961..3549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3450..3469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3703..3720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3879..3906)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3998..4084)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4133..4155
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4147..4164
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4148..4164
                     /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 ccaccatgac cttctctgag
      121 atcctggacc gggtgggcag catgggccac ttccagtttc tgcacgtggc catcctgggc
      181 ctgcctatcc tgaacatggc caaccacaat ctgctgcaga tcttcaccgc agcaacacca
      241 gtgcaccact gcaggccacc tcacaacgca tctaccggac catgggtgct gccaatggga
      301 cctaatggca agcctgagcg gtgtctgaga tttgtgcacc cacccaacgc cagcctgcct
      361 aatgacacac agcgcgccat ggagccatgc ctggatggct gggtgtacaa tagcaccaag
      421 gactccatcg tgacagagtg ggacctggtg tgcaactcca ataagctgaa ggagatggcc
      481 cagtctatct tcatggcagg aatcctgatc ggaggactgg tgctgggcga cctgtctgat
      541 cggtttggcc ggagacctat cctgacctgc agctacctgc tgctggcagc aagcggatcc
      601 ggagcagcct tctccccaac atttcccatc tatatggtgt tccggttcct gtgcggcttc
      661 ggcatctctg gcatcaccct gagcacagtg atcctgaacg tggagtgggt gccaaccagg
      721 atgcgcgcca tcatgtccac cgccctgggc tactgctata cattcggcca gtttatcctg
      781 ccaggactgg catacgcaat cccacagtgg aggtggctgc agctgaccgt gtctatcccc
      841 ttcttcgtgt tcttcctgag ctcctggtgg acacccgagt ccatccgctg gctggtgctg
      901 tctggcaagt ctagcaaggc cctgaagatc ctgaggcgcg tggccgtgtt caacggcaag
      961 aaggaggagg gagagcggct gagcctggag gagctgaagc tgaatctgca gaaggagatc
     1021 agcctggcca aggccaagta taccgcctcc gacctgttca gaatccctat gctgcggaga
     1081 atgacctttt gtctgtccct ggcctggttc gccacaggct ttgcctacta ttctctggcc
     1141 atgggcgtgg aggagtttgg cgtgaatctg tacatcctgc agatcatctt cggcggcgtg
     1201 gacgtgccag ccaagtttat caccatcctg tctctgagct atctgggcag gcacaccaca
     1261 caggcagccg ccctgctgct ggcaggagga gccatcctgg ccctgacatt cgtgcccctg
     1321 gatctgcaga ccgtgcgcac agtgctggcc gtgtttggca agggctgcct gtcctctagc
     1381 ttcagctgtc tgtttctgta cacctccgag ctgtatccca ccgtgatcag gcagacaggc
     1441 atgggcgtgt ccaacctgtg gaccagggtg ggctccatgg tgtctcctct ggtgaagatc
     1501 acaggcgagg tgcagccttt catcccaaat atcatctatg gcatcacagc cctgctggga
     1561 ggaagcgccg ccctgtttct gcccgagacc ctgaaccagc ctctgccaga gacaatcgag
     1621 gatctggaga attggagcct gcgggccaag aagcccaagc aggagcctga ggtggagaag
     1681 gcctcccaga gaatcccact gcagcctcac ggaccaggac tgggctcctc ttaagaccca
     1741 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     1801 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     1861 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     1921 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     1981 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     2041 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     2101 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     2161 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     2221 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     2281 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     2341 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     2401 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     2461 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     2521 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     2581 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     2641 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     2701 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     2761 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     2821 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     2881 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     2941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3601 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3661 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3721 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3781 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3841 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     3901 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     3961 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     4021 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     4081 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     4141 cgacgttgta aaacgacggc cagtcttaag ctcgggccc
//
