LOCUS       sequence-260110-        4146 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 4146)
  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 4146)
  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_SLC1A4
FEATURES             Location/Qualifiers
     source          1..4146
                     /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(2271..2370)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2387..2406)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2388..2406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2411..2427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2411..2427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2529..2548)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2540..3349
                     /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(2607..2626)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3496..4084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3985..4004
                     /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 agagcaacga gacaaatggc tacctggact ccgcccaggc
      721 aggacctgca gcaggaccag gagcacctgg caccgccgcc ggcagggccc ggagatgcgc
      781 cggcttcctg aggcgccagg ccctggtgct gctgaccgtg agcggcgtgc tggcaggagc
      841 aggactggga gccgccctga ggggcctgtc cctgtctcgc acacaggtga cctacctggc
      901 ctttcccggg gagatgttgc tcagaatgct gagaatgatc atcctgccac tggtggtgtg
      961 ctctctggtg agcggagcag ccagcctgga cgcctcctgt ctgggccgcc tgggaggaat
     1021 cgcagtggca tatttcggcc tgaccacact gagcgcctcc gccctggcag tggccctggc
     1081 ctttatcatc aagccaggaa gcggagcaca gacactgcag agctccgacc tgggactgga
     1141 ggattccgga ccacctccag tgccaaagga gaccgtggac tccttcctgg atctggcccg
     1201 gaacctgttt ccttctaatc tggtggtggc cgccttcaga acatacgcca ccgattataa
     1261 ggtggtgacc cagaactcta gctccggcaa tgtgacacac gagaagatcc ctatcggcac
     1321 cgagatcgag ggcatgaaca tcctgggact ggtgctgttc gcactggtgc tgggagtggc
     1381 cctgaagaag ctgggaagcg agggagagga cctgatccgg ttctttaact ctctgaatga
     1441 ggccacaatg gtgctggtga gctggatcat gtggtacgtg cctgtgggca tcatgttcct
     1501 ggtgggctcc aagatcgtgg agatgaagga tatcatcgtg ctggtgacct ccctgggcaa
     1561 gtacatcttt gcctctatcc tgggccacgt gatccacggc ggcatcgtgc tgccactgat
     1621 ctatttcgtg tttacacgga agaatccctt cagatttctg ctgggactgc tggcaccttt
     1681 cgccacagcc tttgccacct gctctagctc cgccaccctg ccatctatga tgaagtgtat
     1741 cgaggagaac aatggcgtgg acaagaggat ctcccgcttc atcctgccca tcggcgccac
     1801 agtgaacatg gatggagcag caatcttcca gtgcgtggca gccgtgttta tcgcccagct
     1861 gaacaatgtg gagctgaatg ccggccagat ctttaccatc ctggtgacag ccaccgcctc
     1921 tagcgtggga gcagcaggag tgccagcagg aggcgtgctg acaatcgcca tcatcctgga
     1981 ggcaatcgga ctgcctaccc acgacctgcc actgatcctg gccgtggact ggatcgtgga
     2041 taggaccaca accgtggtga acgtggaggg cgatgcactg ggagcaggca tcctgcacca
     2101 cctgaatcag aaggccacca agaagggcga gcaggagctg gccgaggtga aggtggaggc
     2161 catcccaaac tgtaagtctg aggaggagac aagccccctg gtgacccacc agaatccagc
     2221 aggaccagtg gcaagcgccc cagagctgga gtccaaggag tctgtgctgg acccagcttt
     2281 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2341 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2401 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2461 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2521 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2581 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2641 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2701 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2761 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2821 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2881 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2941 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3001 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3061 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3121 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3181 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3241 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3301 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3361 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3421 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3601 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4141 catgtt
//
