LOCUS       sequence-263854-        4497 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 4497)
  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 4497)
  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_SLC28A1
FEATURES             Location/Qualifiers
     source          1..4497
                     /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(2622..2721)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2738..2757)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2739..2757)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2762..2778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2762..2778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2880..2899)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2891..3700
                     /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(2958..2977)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3847..4435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4336..4355
                     /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 accatggaga acgaccctag ccggagaagg gagtccatct ctctgacccc
      721 agtggccaag ggcctggaga atatgggcgc cgacttcctg gagtccctgg aggagggaca
      781 gctgccaaga agcgatctgt cccccgccga gatcaggagc tcctggtctg aggcagcacc
      841 aaagcctttt agcagatgga ggaatctgca gcctgccctg agggcacggt ccttctgccg
      901 cgagcacatg cagctgtttc ggtggatcgg aaccggactg ctgtgcacag gactgtctgc
      961 cttcctgctg gtggcctgtc tgctggactt ccagagagcc ctggccctgt ttgtgctgac
     1021 ctgcgtggtg ctgaccttcc tgggccacag actgctgaag aggctgctgg gaccaaagct
     1081 gcgccggttt ctgaagccac agggccaccc cagactgctg ctgtggttca agaggggact
     1141 ggccctggca gcctttctgg gactggtgct gtggctgtcc ctggatacct ctcagagacc
     1201 tgagcagctg gtgtcttttg ccggcatctg cgtgttcgtg gccctgctgt ttgcctgcag
     1261 caagcaccac tgtgccgtgt cttggagggc cgtgagctgg ggactgggac tgcagttcgt
     1321 gctgggactg ctggtcatca gaaccgagcc aggcttcatc gcctttgagt ggctgggcga
     1381 gcagatcagg atctttctga gctacacaaa ggccggctct agcttcgtgt ttggcgaggc
     1441 cctggtgaag gacgtgttcg cctttcaggt gctgcccatc atcgtgttct ttagctgcgt
     1501 gatctccgtg ctgtatcacg tgggcctgat gcagtgggtc atcctgaaga tcgcctggct
     1561 gatgcaggtg accatgggca ccacagccac cgagacactg tccgtggccg gcaacatctt
     1621 cgtgtctcag accgaggcac cactgctgat caggccttac ctggccgaca tgacactgtc
     1681 cgaggtgcac gtggtcatga ccggaggata cgcaacaatc gcaggaagcc tgctgggagc
     1741 ctatatctcc ttcggcatcg atgccacatc cctgatcgca gcatctgtga tggcagcacc
     1801 atgcgccctg gccctgagca agctggtgta ccctgaggtg gaggagtcca agtttagaag
     1861 ggaggagggc gtgaagctga cctatggcga cgcccagaat ctgatcgagg cagcaagcac
     1921 aggagcagca atctccgtga aggtggtggc aaacatcgcc gccaatctga tcgccttcct
     1981 ggccgtgctg gacttcatca acgccgccct gagctggctg ggcgacatgg tggatatcca
     2041 gggcctgtct ttccagctga tctgcagcta tatcctgcgc ccagtggcct ttctgatggg
     2101 agtggcatgg gaggactgtc cagtggtggc agagctgctg ggcatcaagc tgttcctgaa
     2161 tgagtttgtg gcctaccagg acctgagcaa gtataagcag cgccggctgg caggagcaga
     2221 ggagtgggtg ggcgatcgca agcagtggat ctccgtgcgg gccgaggtgc tgaccacatt
     2281 cgccctgtgc ggcttcgcca acttttcctc tatcggcatc atgctgggag gactgaccag
     2341 catggtgcct cagcgcaagt ctgacttcag ccagatcgtg ctgcgggccc tgtttacagg
     2401 cgcctgcgtg tctctggtga acgcctgtat ggcaggaatc ctgtacatgc caaggggagc
     2461 cgaggtggat tgcatgtccc tgctgaatac cacactgagc tcctctagct tcgagatcta
     2521 tcagtgctgt cgcgaggcct tccagtccgt gaaccctgag ttttctccag aggccctgga
     2581 taactgctgt cggttttaca atcacaccat ctgtgcccag gacccagctt tcttgtacaa
     2641 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2701 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2761 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2821 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2881 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2941 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     3001 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3061 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3121 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3181 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3241 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3301 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3361 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3421 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3481 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3541 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3601 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3661 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3721 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3781 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3841 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3901 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3961 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4021 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4081 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4141 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4201 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4261 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4321 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4381 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4441 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
