LOCUS       sequence-333106-        4284 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 4284)
  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 4284)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      784..793
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1873..1972)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1989..2008)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1990..2008)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2013..2029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2013..2029)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2131..2150)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2142..2951
                     /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(2209..2228)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3098..3686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3587..3606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3840..3857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4016..4043)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4135..4221)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4270..4284,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4284,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacac attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccaccatg ggcagccgcc acttcgaggg catctacgac cacgtgggcc
      181 acttcggccg gtttcagaga gtgctgtatt tcatctgcgc ctttcagaac atctcttgtg
      241 gcatccacta cctggccagc gtgttcatgg gagtgacccc tcaccacgtg tgcaggccac
      301 ctggaaacgt gagccaggtg gtgtttcaca accactccaa ttggtctctg gaggacacag
      361 gcgccctgct gagctccgga cagaaggatt acgtgacagt tcaactgcag aatggtgaga
      421 tctgggagct gagcaggtgt tcccgcaaca agcgggagaa tacatctagc ctgggctacg
      481 agtataccgg ctctaagaag gagttccctt gcgtggacgg ctacatctat gatcagaaca
      541 cctggaagag caccgccgtg acacagtgga acctggtgtg cgatcgcaag tggctggcca
      601 tgctgatcca gccactgttc atgtttggcg tgctgctggg ctccgtgaca ttcggctatt
      661 tttctgacag gctgggccgg agagtggtgc tgtgggccac ctcctctagc atgttcctgt
      721 ttggcatcgc cgccgccttc gccgtggatt actatacctt catggccgcc cggttctttc
      781 tggccatggt ggccagcggc tacctggtgg tgggcttcgt gtatgtgatg gagtttatcg
      841 gcatgaagtc tagaacctgg gccagcgtgc acctgcactc cttctttgca gtgggcaccc
      901 tgctggtggc cctgacaggc tacctggtga ggacctggtg gctgtatcag atgatcctgt
      961 ccaccgtgac agtgccattc atcctgtgct gttgggtgct gccagagaca cccttttggc
     1021 tgctgtctga gggccgctac gaggaggccc agaagatcgt ggacatcatg gccaagtgga
     1081 accgggcctc ctcttgcaag ctgtctgagc tgctgagcct ggatctgcag ggccccgtga
     1141 gcaattctcc aaccgaggtg cagaagcaca acctgagcta cctgttctat aattggtcca
     1201 tcacaaagag aaccctgaca gtgtggctga tctggtttac cggctccctg ggcttctata
     1261 gcttttccct gaactctgtg aatctgggcg gcaacgagta cctgaatctg ttcctgctgg
     1321 gcgtggtgga gatccccgcc tataccttcg tgtgcatcgc catggacaaa gtgggcaggc
     1381 gcaccgtgct ggcatactcc ctgttctgct ctgccctggc ctgtggcgtg gtcatggtca
     1441 tcccacagaa gcactatatc ctgggcgtgg tgacagccat ggtgggcaag ttcgccatcg
     1501 gcgccgcctt tggcctgatc tacctgtata cagccgagct gtaccccacc atcgtgagaa
     1561 gcctggccgt gggctctggc agcatggtgt gcaggctggc ctctatcctg gcccccttta
     1621 gcgtggacct gagctccatc tggatcttca tccctcagct gtttgtggga accatggcac
     1681 tgctgtccgg cgtgctgaca ctgaagctgc ccgagaccct gggcaagaga ctggccacca
     1741 catgggagga ggccgccaag ctggagagcg agaatgagtc caagtctagc aagctgctgc
     1801 tgaccacaaa caatagcggc ctggagaaga ccgaggccat cacacctagg gattccggcc
     1861 tgggcgagta agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
     1921 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     1981 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     2041 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     2101 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     2161 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     2221 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     2281 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2341 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2401 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     2461 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     2521 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2581 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2641 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     2701 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     2761 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     2821 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     2881 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     2941 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     3001 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     3061 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3121 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3181 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3241 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3301 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3361 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3421 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3481 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3541 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3601 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3661 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3721 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3781 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3841 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3901 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3961 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
     4021 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
     4081 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
     4141 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
     4201 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
     4261 ggatgttttc ccagtcacga cgtt
//
