LOCUS       sequence-259085-        4167 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 4167)
  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 4167)
  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_SLC16A2
FEATURES             Location/Qualifiers
     source          1..4167
                     /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(2292..2391)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2408..2427)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2409..2427)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2432..2448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2432..2448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2550..2569)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2561..3370
                     /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(2628..2647)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3517..4105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4006..4025
                     /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 accatggcac tgcagtctca ggcaagcgag gaggcaaagg gaccatggca
      721 ggaggcagac caggagcagc aggagcctgt gggaagccca gagcctgagt ccgagccaga
      781 gccagagcct gagccagagc cagtgcctgt gccacctcca gagccacagc cagagcctca
      841 gccactgcca gatcctgcac cactgccaga gctggagttc gagtctgaga gggtgcacga
      901 gcctgagcca accccaacag tggagaccag gggaacagca aggggatttc agccacctga
      961 gggaggattc ggatgggtgg tggtgtttgc cgccacctgg tgcaacggct ccatcttcgg
     1021 catccacaat tctgtgggca tcctgtacag catgctgctg gaggaggaga aggagaagaa
     1081 ccgccaggtg gagtttcagg cagcatgggt gggcgcactt gctatgggta tgatattttt
     1141 ctgctcccct atcgtgtcta tcttcaccga ccggctgggc tgtagaatca ccgcaacagc
     1201 aggagcagca gtggccttca tcggcctgca caccagctcc tttacatcta gcctgagcct
     1261 gcggtacttc acatatggca tcctgtttgg ctgcggctgt tccttcgcct ttcagccttc
     1321 tctggtcatc ctgggccact acttccagcg gagactggga ctggcaaacg gagtggtgtc
     1381 cgccggctcc tctatctttt ccatgtcttt cccatttctg atccgcatgc tgggcgacaa
     1441 gatcaagctg gcccagacct ttcaggtgct gtccacattc atgtttgtgc tgatgctgct
     1501 gtctctgacc tatcggcccc tgctgcctag ctcccaggat accccaagca agcggggcgt
     1561 gagaacactg caccagaggt tcctggccca gctgcgcaag tacttcaata tgcgggtgtt
     1621 taggcagcgc acatatagaa tctgggcctt cggcatcgca gcagccgccc tgggatactt
     1681 tgtgccctat gtgcacctga tgaagtacgt ggaggaggag ttctccgaga tcaaggagac
     1741 ctgggtgctg ctggtgtgca tcggagccac aagcggactg ggcagactgg tgtccggaca
     1801 catcagcgac tccatcccag gcctgaagaa gatctatctg caggtgctgt cttttctgct
     1861 gctgggcctg atgagcatga tgatccccct gtgcagggac ttcggcggcc tgatcgtggt
     1921 gtgcctgttt ctgggcctgt gcgatggctt ctttatcacc atcatggcac caatcgcctt
     1981 cgagctggtg ggaccaatgc aggcaagcca ggccatcggc tatctgctgg gcatgatggc
     2041 cctgcccatg atcgcaggac caccaatcgc aggactgctg aggaattgtt ttggcgatta
     2101 ccacgtggcc ttctattttg caggagtgcc tccaatcatc ggagccgtga tcctgttctt
     2161 tgtgcctctg atgcaccaga gaatgttcaa gaaggagcag agggactcta gcaaggataa
     2221 gatgctggcc cctgacccag atccaaacgg agagctgctg cctggaagcc ctaatccaga
     2281 ggagccaatc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2341 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2401 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2461 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2521 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2581 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2641 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2701 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2761 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2821 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2881 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2941 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3001 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3061 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3121 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3181 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3241 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3301 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3361 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3421 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3481 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3541 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3601 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3661 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3721 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3781 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3841 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3901 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3961 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4021 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4081 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4141 cttttgctgg ccttttgctc acatgtt
//
