LOCUS       sequence-261015-        4281 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 4281)
  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 4281)
  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_SLC22A16
FEATURES             Location/Qualifiers
     source          1..4281
                     /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"
     regulatory      1320..1329
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2406..2505)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2522..2541)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2523..2541)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2546..2562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2546..2562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2664..2683)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2675..3484
                     /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(2742..2761)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3631..4219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4120..4139
                     /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 accatgggca gccgccactt cgagggcatc tacgaccacg tgggccactt
      721 cggccggttt cagagagtgc tgtatttcat ctgcgccttt cagaacatct cttgtggcat
      781 ccactacctg gccagcgtgt tcatgggagt gacccctcac cacgtgtgca ggccacctgg
      841 aaacgtgagc caggtggtgt ttcacaacca ctccaattgg tctctggagg acacaggcgc
      901 cctgctgagc tccggacaga aggattacgt gacagttcaa ctgcagaatg gtgagatctg
      961 ggagctgagc aggtgttccc gcaacaagcg ggagaataca tctagcctgg gctacgagta
     1021 taccggctct aagaaggagt tcccttgcgt ggacggctac atctatgatc agaacacctg
     1081 gaagagcacc gccgtgacac agtggaacct ggtgtgcgat cgcaagtggc tggccatgct
     1141 gatccagcca ctgttcatgt ttggcgtgct gctgggctcc gtgacattcg gctatttttc
     1201 tgacaggctg ggccggagag tggtgctgtg ggccacctcc tctagcatgt tcctgtttgg
     1261 catcgccgcc gccttcgccg tggattacta taccttcatg gccgcccggt tctttctggc
     1321 catggtggcc agcggctacc tggtggtggg cttcgtgtat gtgatggagt ttatcggcat
     1381 gaagtctaga acctgggcca gcgtgcacct gcactccttc tttgcagtgg gcaccctgct
     1441 ggtggccctg acaggctacc tggtgaggac ctggtggctg tatcagatga tcctgtccac
     1501 cgtgacagtg ccattcatcc tgtgctgttg ggtgctgcca gagacaccct tttggctgct
     1561 gtctgagggc cgctacgagg aggcccagaa gatcgtggac atcatggcca agtggaaccg
     1621 ggcctcctct tgcaagctgt ctgagctgct gagcctggat ctgcagggcc ccgtgagcaa
     1681 ttctccaacc gaggtgcaga agcacaacct gagctacctg ttctataatt ggtccatcac
     1741 aaagagaacc ctgacagtgt ggctgatctg gtttaccggc tccctgggct tctatagctt
     1801 ttccctgaac tctgtgaatc tgggcggcaa cgagtacctg aatctgttcc tgctgggcgt
     1861 ggtggagatc cccgcctata ccttcgtgtg catcgccatg gacaaagtgg gcaggcgcac
     1921 cgtgctggca tactccctgt tctgctctgc cctggcctgt ggcgtggtca tggtcatccc
     1981 acagaagcac tatatcctgg gcgtggtgac agccatggtg ggcaagttcg ccatcggcgc
     2041 cgcctttggc ctgatctacc tgtatacagc cgagctgtac cccaccatcg tgagaagcct
     2101 ggccgtgggc tctggcagca tggtgtgcag gctggcctct atcctggccc cctttagcgt
     2161 ggacctgagc tccatctgga tcttcatccc tcagctgttt gtgggaacca tggcactgct
     2221 gtccggcgtg ctgacactga agctgcccga gaccctgggc aagagactgg ccaccacatg
     2281 ggaggaggcc gccaagctgg agagcgagaa tgagtccaag tctagcaagc tgctgctgac
     2341 cacaaacaat agcggcctgg agaagaccga ggccatcaca cctagggatt ccggcctggg
     2401 cgaggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2461 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2521 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2581 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2641 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2701 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2761 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2821 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2881 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2941 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3001 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3061 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3121 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3181 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3241 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3301 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3361 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3421 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3481 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3541 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3601 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3661 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3721 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3781 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3841 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3901 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3961 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4021 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4081 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4141 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4201 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4261 ctggcctttt gctcacatgt t
//
