LOCUS       sequence-259576-        4254 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 4254)
  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 4254)
  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_SLC13A5
FEATURES             Location/Qualifiers
     source          1..4254
                     /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(2379..2478)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2495..2514)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2496..2514)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2519..2535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2519..2535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2637..2656)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2648..3457
                     /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(2715..2734)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3604..4192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4093..4112
                     /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 accatggcct ctgccctgag ctacgtgtcc aagtttaagt ccttcgtgat
      721 cctgtttgtg accccactgc tgctgctgcc cctggtcatc ctgatgccag ccaagttcgt
      781 gaggtgcgcc tacgtgatca tcctgatggc catctattgg tgtaccgaag tgatccccct
      841 ggccgtgaca agcctgatgc ctgtgctgct gtttccactg ttccagatcc tggactcccg
      901 ccaggtgtgc gtgcagtata tgaaggatac caacatgctg tttctgggag gactgatcgt
      961 ggcagtggca gtggagaggt ggaatctgca caagaggatc gccctgcgca cactgctgtg
     1021 ggtgggagca aagcctgcaa ggctgatgct gggcttcatg ggcgtgaccg ccctgctgtc
     1081 tatgtggatc agcaacacag ccaccacagc catgatggtg ccaatcgtgg aggccatcct
     1141 gcagcagatg gaggccacat ccgccgcaac cgaggcagga ctggagctgg tggacaaggg
     1201 caaggccaag gagctgccag gctctcaggt catcttcgag ggacccaccc tgggacagca
     1261 ggaggatcag gagcggaagc ggctgtgcaa ggccatgaca ctgtgcatct gttacgcagc
     1321 atccatcgga ggaacagcca ccctgacagg aaccggacct aacgtggtgc tgctgggaca
     1381 gatgaatgag ctgtttccag acagcaagga tctggtgaac ttcgcctcct ggttcgcctt
     1441 tgccttccca aatatgctgg tcatgctgct gtttgcctgg ctgtggctgc agttcgtgta
     1501 catgcggttt aacttcaaga agtcctgggg ctgcggcctg gagtctaaga agaatgagaa
     1561 ggccgccctg aaggtgctgc aggaggagta tcggaagctg ggccccctgt ctttcgccga
     1621 gatcaacgtg ctgatctgtt tctttctgct ggtcatcctg tggtttagca gagaccctgg
     1681 cttcatgcca ggatggctga ccgtggcatg ggtggaggga gagacaaagt acgtgagcga
     1741 tgccaccgtg gccatctttg tggccacact gctgttcatc gtgccctctc agaagcctaa
     1801 gtttaatttc cggagccaga cagaggagga gagaaagacc cccttttatc cccctccact
     1861 gctggactgg aaggtgaccc aggagaaggt gccttggggc atcgtgctgc tgctgggagg
     1921 aggattcgca ctggcaaagg gatctgaggc aagcggactg tccgtgtgga tgggcaagca
     1981 gatggagcct ctgcacgcag tgccacctgc agccatcacc ctgatcctgt ccctgctggt
     2041 ggccgtgttt acagagtgca cctctaacgt ggccaccaca accctgtttc tgcccatctt
     2101 cgccagcatg tcccggtcta tcggcctgaa tcccctgtac atcatgctgc cttgtacact
     2161 gagcgcctcc tttgccttca tgctgcctgt ggccacccca cccaacgcca tcgtgttcac
     2221 atatggccac ctgaaggtgg ccgacatggt gaagaccggc gtgatcatga acatcatcgg
     2281 cgtgttttgc gtgttcctgg ccgtgaatac atggggcaga gccatctttg acctggatca
     2341 cttccccgat tgggccaatg tgacacacat cgagaccgac ccagctttct tgtacaaagt
     2401 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2461 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2521 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2581 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2641 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2701 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2761 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2821 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2881 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2941 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     3001 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     3061 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     3121 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3181 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3241 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3301 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3361 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3421 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3481 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3541 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3601 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3661 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3721 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3781 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3841 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3901 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3961 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4021 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4081 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4141 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4201 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
