LOCUS       sequence-259274-        4335 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 4335)
  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 4335)
  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_SLC13A1
FEATURES             Location/Qualifiers
     source          1..4335
                     /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"
     regulatory      1103..1112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2460..2559)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2576..2595)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2577..2595)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2600..2616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2600..2616)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2718..2737)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2729..3538
                     /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(2796..2815)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3685..4273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4174..4193
                     /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 accatgaagt tctttagcta catcctggtg tatcggagat tcctgtttgt
      721 ggtgtttaca gtgctggtgc tgctgccact gcccatcgtg ctgcacacaa aggaggccga
      781 gtgcgcctac accctgtttg tggtggccac attctggctg accgaggccc tgccactgtc
      841 tgtgaccgcc ctgctgccaa gcctgatgct gcctatgttc ggcatcatgc cttccaagaa
      901 ggtggcctct gcctacttta aggacttcca cctgctgctg atcggcgtga tctgtctggc
      961 cacctccatc gagaagtgga acctgcacaa gagaatcgcc ctgaagatgg tcatgatggt
     1021 gggcgtgaat cctgcctggc tgacactggg ctttatgagc tccaccgcct tcctgtctat
     1081 gtggctgtct aacaccagca cagccgccat ggtcatgcca atcgccgagg ccgtggtgca
     1141 gcagatcatc aatgccgagg ccgaggtgga ggccacacag atgacctact tcaacggctc
     1201 caccaatcac ggcctggaga tcgacgagtc tgtgaacggc cacgagatca atgagcggaa
     1261 ggagaagaca aagcctgtgc caggctataa caatgatacc ggcaagatct ctagcaaggt
     1321 ggagctggag aagaacagcg gcatgaggac aaagtaccgc accaagaagg gccacgtgac
     1381 acggaagctg acctgcctgt gcatcgccta ttcctctacc atcggcggcc tgaccacaat
     1441 caccggcaca tccaccaacc tgatctttgc cgagtacttc aatacacggt atcctgactg
     1501 cagatgtctg aactttggca gctggttcac cttttccttc ccagccgccc tgatcatcct
     1561 gctgctgtcc tggatctggc tgcagtggct gtttctgggc tttaatttca aggagatgtt
     1621 caagtgcggc aagacaaaga ccgtgcagca gaaggcctgt gccgaagtga tcaagcagga
     1681 gtaccagaag ctgggcccca tcaggtatca ggagatcgtg accctggtgc tgttcatcat
     1741 catggccctg ctgtggtttt ctagggaccc aggattcgtg cctggatgga gcgccctgtt
     1801 ttccgagtac cctggcttcg ccacagatag caccgtggcc ctgctgatcg gcctgctgtt
     1861 ctttctgatc cccgccaaga cactgaccaa gaccacacct acaggcgaga tcgtggcctt
     1921 tgattatagc ccactgatca cctggaagga gtttcagtcc ttcatgccat gggacatcgc
     1981 aatcctggtg ggaggaggat tcgcactggc agatggatgc gaggagtccg gactgtctaa
     2041 gtggatcggc aataagctgt ctccactggg aagcctgcca gcatggctga tcatcctgat
     2101 cagctccctg atggtgacat ccctgaccga ggtggcctct aaccccgcca caatcaccct
     2161 gttcctgcca atcctgagcc cactggcaga ggcaatccac gtgaatcctc tgtacatcct
     2221 gatcccatct acactgtgca ccagctttgc cttcctgctg ccagtggcaa acccacctaa
     2281 tgccatcgtg ttttcctatg gccacctgaa agtgatcgac atggtgaagg caggactggg
     2341 agtgaacatc gtgggagtgg ccgtggtcat gctgggcatc tgtacatgga tcgtgccaat
     2401 gttcgatctg tacacctatc ctagctgggc cccagccatg tccaatgaga ccatgcccga
     2461 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2521 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2581 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2641 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2701 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2761 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2821 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2881 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2941 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     3001 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     3061 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3121 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3181 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3241 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3301 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3361 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3421 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3481 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3541 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3601 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3661 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3721 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3781 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3841 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3901 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3961 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4021 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4081 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4141 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4201 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4261 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4321 ttttgctcac atgtt
//
