LOCUS       sequence-260033-        4173 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 4173)
  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 4173)
  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_SLC1A5
FEATURES             Location/Qualifiers
     source          1..4173
                     /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      672..681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1466..1475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2298..2397)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2414..2433)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2415..2433)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2438..2454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2438..2454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2556..2575)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2567..3376
                     /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(2634..2653)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3523..4111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4012..4031
                     /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 accatggtgg ccgacccccc tagagattcc aagggactgg cagcagcaga
      721 gcctacagcc aacggcggcc tggccctggc cagcatcgag gaccagggag cagcagcagg
      781 aggatactgc ggctccaggg atcaggtgcg gagatgtctg cgcgccaatc tgctggtgct
      841 gctgaccgtg gtggccgtgg tggccggcgt ggccctgggc ctgggcgtgt ctggagcagg
      901 aggcgccctg gccctgggac ctgagcggct gagcgccttc gtgtttccag gagagctgct
      961 gctgaggctg ctgaggatga tcatcctgcc tctggtggtg tgcagcctga tcggaggagc
     1021 agcatccctg gacccaggcg ccctgggcag actgggcgcc tgggccctgc tgttctttct
     1081 ggtgaccaca ctgctggcca gcgccctggg agtgggactg gccctggccc tgcagccagg
     1141 cgccgcctcc gccgccatca acgcctctgt gggcgccgcc ggcagcgccg agaatgcccc
     1201 ctccaaggaa gtcctggact cttttcttga tctggccagg aacatctttc ctagcaatct
     1261 ggtgtccgcc gccttccgca gctactccac cacatatgag gagcggaaca tcaccggcac
     1321 aagagtgaag gtgccagtgg gacaggaggt ggagggcatg aatatcctgg gcctggtggt
     1381 gttcgccatc gtgtttggag tggccctgag gaagctggga ccagagggag agctgctgat
     1441 cagattcttt aactctttca atgaggccac catggtgctg gtgagctgga tcatgtggta
     1501 cgcccctgtg ggcatcatgt tcctggtggc cggcaagatc gtggagatgg aggacgtggg
     1561 actgctgttt gcaaggctgg gcaagtatat cctgtgctgt ctgctgggac acgcaatcca
     1621 cggactgctg gtgctgcccc tgatctactt cctgtttaca aggaagaacc cctatcgctt
     1681 cctgtggggc atcgtgaccc ctctggccac agcctttggc accagctcct ctagcgccac
     1741 cctgcctctg atgatgaagt gcgtggagga gaacaatggc gtggccaagc acatctcccg
     1801 gttcatcctg ccaatcggcg ccacagtgaa tatggacgga gccgccctgt tccagtgcgt
     1861 ggcagccgtg tttatcgccc agctgtccca gcagtctctg gattttgtga agatcatcac
     1921 aatcctggtg accgccacag cctcctctgt gggagcagca ggaatcccag caggaggcgt
     1981 gctgaccctg gcaatcatcc tggaggccgt gaacctgccc gtggatcaca tctccctgat
     2041 cctggccgtg gactggctgg tggataggtc ttgtaccgtg ctgaacgtgg agggcgacgc
     2101 actgggagca ggactgctgc agaattacgt ggatcggaca gagtctagaa gcaccgagcc
     2161 agagctgatc caggtgaaga gcgagctgcc actggaccca ctgccagtgc ctacagagga
     2221 gggcaatcca ctgctgaagc actatagggg accagcaggc gatgcaaccg tggcatctga
     2281 gaaggagagc gtgatggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2341 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2401 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2461 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2521 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2581 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2641 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2701 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2761 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2821 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2881 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2941 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3001 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3061 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3121 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3181 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3241 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3301 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3361 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3421 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3481 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3541 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3601 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3661 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3721 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3781 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3841 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3901 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3961 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4021 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4081 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4141 cctggccttt tgctggcctt ttgctcacat gtt
//
