LOCUS       sequence-258687-        4476 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 4476)
  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 4476)
  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_SLC6A14
FEATURES             Location/Qualifiers
     source          1..4476
                     /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(2601..2700)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2717..2736)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2718..2736)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2741..2757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2741..2757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2859..2878)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2870..3679
                     /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(2937..2956)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3826..4414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4315..4334
                     /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 accatggaca agctgaagtg cccttctttc tttaagtgta gggagaagga
      721 gaaggtgagc gccagctccg agaacttcca cgtgggcgag aacgacgaga atcaggatcg
      781 cggcaattgg tccaagaagt ctgattacct gctgtctatg atcggctatg ccgtgggcct
      841 gggcaacgtg tggaggttcc catacctgac atatagcaat ggcggcggcg cctttctgat
      901 cccatacgca atcatgctgg ccctggcagg actgcccctg ttctttctgg agtgctccct
      961 gggacagttt gcaagcctgg gaccagtgtc cgtgtggagg atcctgccac tgttccaggg
     1021 agtgggcatc acaatggtgc tgatctccat ctttgtgacc atctactata acgtgatcat
     1081 cgcctattct ctgtactata tgtttgccag cttccagtcc gagctgcctt ggaagaactg
     1141 ctctagctgg agcgacaaga attgttcccg gtctccaatc gtgacacact gcaacgtgag
     1201 caccgtgaat aagggcatcc aggagatcat ccagatgaac aagtcctggg tggatatcaa
     1261 caatttcaca tgtatcaatg gcagcgagat ctaccagcct ggccagctgc catccgagca
     1321 gtattggaac aaggtggccc tgcagcggtc ctctggcatg aatgagaccg gcgtgatcgt
     1381 gtggtacctg gccctgtgcc tgctgctggc atggctgatc gtgggagccg ccctgtttaa
     1441 gggcatcaag agctccggca aggtggtgta cttcacagcc ctgtttccct atgtggtgct
     1501 gctgatcctg ctggtgagag gagccaccct ggagggagca tctaagggaa tcagctacta
     1561 tatcggcgcc cagtccaact tcacaaagct gaaggaggca gaagtgtgga aggacgcagc
     1621 aacccagatc ttttacagcc tgtccgtggc atggggagga ctggtggccc tgtctagcta
     1681 caacaagttc aagaacaact gttttagcga tgccatcgtg gtgtgcctga caaattgtct
     1741 gacctccgtg tttgccggct tcgccatctt ttctatcctg ggccacatgg cccacatctc
     1801 tggcaaggag gtgagccagg tggtgaagtc cggcttcgac ctggccttta tcgcctaccc
     1861 tgaggccctg gcacagctgc caggaggacc attctggtcc atcctgttct ttttcatgct
     1921 gctgacactg ggcctggact cccagttcgc ctctatcgag accatcacca caaccatcca
     1981 ggatctgttt cccaaagtga tgaagaagat gagggtgcct atcacactgg gctgctgtct
     2041 ggtgctgttc ctgctgggac tggtgtgcgt gacccaggca ggcatctact gggtgcacct
     2101 gatcgaccac ttttgtgcag gatggggaat cctgatcgca gcaatcctgg agctggtggg
     2161 catcatctgg atctatggcg gcaaccggtt catcgaggat acagagatga tgatcggcgc
     2221 caagcggtgg atcttctggc tgtggtggag agcctgctgg tttgtgatca cccccatcct
     2281 gctgatcgcc atcttcatct ggagcctggt gcagtttcac agacctaatt acggcgccat
     2341 cccatatcct gactggggag tggccctggg atggtgcatg atcgtgttct gtatcatctg
     2401 gatcccaatc atggccatca tcaagatcat ccaggccaag ggcaacatct ttcagaggct
     2461 gatctcttgc tgtcgccccg ccagcaattg gggaccttac ctggagcagc acaggggaga
     2521 gagatataag gacatggtgg atcccaagaa ggaggccgat cacgagatcc ctaccgtgtc
     2581 tggcagccgc aagccagagg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2641 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2701 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2761 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2821 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2881 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2941 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     3001 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3061 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3121 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3181 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3241 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3301 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3361 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3421 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3481 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3541 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3601 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3661 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3721 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3781 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3841 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3901 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3961 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4021 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4081 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4141 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4201 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4261 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4321 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4381 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4441 gttcctggcc ttttgctggc cttttgctca catgtt
//
