LOCUS       sequence-259328-        4455 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 4455)
  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 4455)
  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_SLC6A8
FEATURES             Location/Qualifiers
     source          1..4455
                     /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(2580..2679)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2696..2715)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2697..2715)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2720..2736)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2720..2736)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2838..2857)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2849..3658
                     /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(2916..2935)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3805..4393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4294..4313
                     /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 accatggcca agaagtctgc cgagaatggc atctactccg tgtctggcga
      721 cgagaagaag ggccccctga tcgccccagg ccccgacggc gccccagcca agggcgatgg
      781 ccccgtgggc ctgggcacac ctggaggccg gctggcagtg ccacctaggg agacatggac
      841 cagacagatg gacttcatca tgagctgcgt gggattcgca gtgggactgg gaaacgtgtg
      901 gagatttcct tacctgtgct ataagaacgg cggcggcgtg ttcctgatcc catacgtgct
      961 gatcgccctg gtgggaggaa tccccatctt ctttctggag atctctctgg gccagttcat
     1021 gaaggccggc agcatcaacg tgtggaacat ctgtccactg tttaagggcc tgggctacgc
     1081 ctccatggtc atcgtgttct actgcaatac atactatatc atggtgctgg cctggggctt
     1141 ttactatctg gtgaagtctt tcaccacaac cctgccttgg gcaacctgcg gacacacatg
     1201 gaacacccca gactgcgtgg agatctttag gcacgaggat tgcgccaatg cctccctggc
     1261 caacctgacc tgtgaccagc tggccgatcg gagatctcca gtgatcgagt tctgggagaa
     1321 taaggtgctg aggctgagcg gaggactgga ggtgccaggc gccctgaact gggaggtgac
     1381 actgtgcctg ctggcctgtt gggtgctggt gtacttttgc gtgtggaagg gcgtgaagtc
     1441 caccggcaag atcgtgtact ttacagccac cttcccctat gtggtgctgg tggtgctgct
     1501 ggtgcggggc gtgctgctgc ctggcgccct ggacggcatc atctactatc tgaagccaga
     1561 ttggtccaag ctgggatctc cacaagtgtg gatcgacgca ggcacacaga tcttcttttc
     1621 ttatgccatc ggactgggcg ccctgaccgc cctgggaagc tacaataggt tcaacaataa
     1681 ctgctataag gatgccatca tcctggccct gatcaactct ggcacaagct tctttgccgg
     1741 ctttgtggtg ttcagcatcc tgggctttat ggccgccgag cagggcgtgc acatcagcaa
     1801 ggtggcagag tccggacctg gactggcctt catcgcatac ccaagggcag tgacactgat
     1861 gccagtggca cctctgtggg ccgccctgtt ctttttcatg ctgctgctgc tgggcctgga
     1921 cagccagttt gtgggcgtgg agggcttcat caccggactg ctggatctgc tgccagcatc
     1981 ctactacttc cggttccagc gcgagatctc tgtggccctg tgctgtgccc tgtgctttgt
     2041 gatcgacctg tccatggtga ccgatggcgg catgtacgtg tttcagctgt tcgactacta
     2101 tagcgcctcc ggaacaaccc tgctgtggca ggccttttgg gagtgcgtgg tggtggcatg
     2161 ggtgtacgga gcagataggt tcatggacga tatcgcctgc atgatcggct atcgcccttg
     2221 cccatggatg aagtggtgtt ggtccttttt cacacctctg gtgtgcatgg gcatctttat
     2281 cttcaacgtg gtgtactatg agcccctggt gtacaataac acctacgtgt atccttggtg
     2341 gggagaggca atgggatggg ccttcgccct gagctccatg ctgtgcgtgc ctctgcacct
     2401 gctgggatgt ctgctgaggg caaagggaac aatggccgag agatggcagc acctgaccca
     2461 gccaatctgg ggcctgcacc acctggagta tagggcacag gacgcagatg tgcgcggact
     2521 gacaaccctg acccccgtga gcgagtctag caaggtggtg gtggtggagt ccgtgatgga
     2581 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2641 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2701 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2761 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2821 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2881 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2941 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     3001 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     3061 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     3121 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     3181 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3241 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3301 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3361 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3421 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3481 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3541 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3601 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3661 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3721 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3781 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3841 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3901 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3961 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4021 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4081 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4141 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4201 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4261 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4321 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4381 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4441 ttttgctcac atgtt
//
