LOCUS       sequence-261208-        4458 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 4458)
  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 4458)
  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_SLC6A7
FEATURES             Location/Qualifiers
     source          1..4458
                     /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(2583..2682)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2699..2718)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2700..2718)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2723..2739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2723..2739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2841..2860)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2852..3661
                     /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(2919..2938)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3808..4396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4297..4316
                     /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 accatgaaga agctgcaggg agcacacctg aggaagcctg tgacaccaga
      721 cctgctgatg accccttctg atcagggcga cgtggatctg gacgtggatt ttgccgccca
      781 cagaggcaac tggacaggca agctggactt cctgctgagc tgcatcggct actgcgtggg
      841 cctgggcaac gtgtggaggt tcccataccg cgcctatacc aatggcggcg gcgcctttct
      901 ggtgccctac ttcctgatgc tggccatctg cggcatccct ctgttctttc tggagctgtc
      961 tctgggacag tttagctccc tgggaccact ggccgtgtgg aagatcagcc ctctgttcaa
     1021 gggagcagga gcagcaatgc tgctgatcgt gggcctggtg gccatctact ataacatgat
     1081 catcgcctac gtgctgtttt atctgttcgc cagcctgaca tccgacctgc cctgggagca
     1141 ctgcggcaac tggtggaata ccgagctgtg cctggagcac cgggtgagca aggatggcaa
     1201 cggcgccctg cctctgaatc tgacctgcac agtgagccca tccgaggagt actggtccag
     1261 atatgtgctg cacatccagg gaagccaggg aatcggatcc ccaggcgaga tcagatggaa
     1321 tctgtgcctg tgcctgctgc tggcctgggt catcgtgttc ctgtgcatcc tgaagggcgt
     1381 gaagtctagc ggcaaggtgg tgtactttac cgccacattc ccctatctga tcctgctgat
     1441 gctgctggtg cggggagtga cactgccagg agcctggaag ggcatccagt tttacctgac
     1501 cccccagttc caccacctgc tgtcctctaa agtgtggatc gaggccgccc tgcagatctt
     1561 ttatagcctg ggagtgggat ttggaggact gctgacattc gcctcctaca acacctttca
     1621 ccagaatatc tatagagaca cattcatcgt gaccctgggc aatgccatca cctccatcct
     1681 ggccggcttt gccatcttct ctgtgctggg ctacatgagc caggagctgg gcgtgcctgt
     1741 ggatcaggtg gcaaaggcag gaccaggact ggccttcgtg gtgtatcccc aggccatgac
     1801 aatgctgccc ctgtctcctt tttggagctt cctgttcttt ttcatgctgc tgaccctggg
     1861 cctggactcc cagtttgcct tcctggagac catcgtgaca gccgtgaccg atgagtttcc
     1921 atactatctg aggcccaaga aggccgtgtt cagcggcctg atctgcgtgg ccatgtacct
     1981 gatgggcctg atcctgacca cagacggcgg catgtactgg ctggtgctgc tggacgatta
     2041 ttctgccagc tttggcctga tggtggtggt catcaccaca tgcctggccg tgaccagggt
     2101 gtacggcatc cagcggttct gtagagatat ccacatgatg ctgggcttca agcctggcct
     2161 gtattttcgc gcatgctggc tgttcctgtc cccagccacc ctgctggccc tgatggtgta
     2221 ctctatcgtg aagtatcagc cctccgagta cggctcttat aggtttccac cttgggcaga
     2281 gctgctggga atcctgatgg gactgctgag ctgtctgatg atccctgcag gaatgctggt
     2341 ggccgtgctg agggaggagg gatccctgtg ggagcggctg cagcaggcct ctagacctgc
     2401 catggactgg ggcccaagcc tggaggagaa ccgcacagga atgtacgtgg ccaccctggc
     2461 aggatcccag tctccaaagc cactgatggt gcacatgagg aagtatggcg gcatcacatc
     2521 cttcgagaat accgccatcg aggtggatcg cgagatcgcc gaggaggagg agtctatgat
     2581 ggacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2641 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2701 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2761 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2821 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2881 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2941 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     3001 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     3061 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     3121 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     3181 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     3241 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3301 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3361 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3421 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3481 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3541 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3601 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3661 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3721 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3781 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4381 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4441 gccttttgct cacatgtt
//
