LOCUS       sequence-259092-        4176 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 4176)
  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 4176)
  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_SLC1A3
FEATURES             Location/Qualifiers
     source          1..4176
                     /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(2301..2400)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2417..2436)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2418..2436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2441..2457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2441..2457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2559..2578)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2570..3379
                     /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(2637..2656)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3526..4114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4015..4034
                     /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 accatgacca agtctaacgg cgaggagcca aagatgggcg gccggatgga
      721 gagattccag cagggcgtgc ggaagagaac cctgctggcc aagaagaagg tgcagaacat
      781 cacaaaggag gacgtgaagt cctacctgtt ccggaatgcc tttgtgctgc tgaccgtgac
      841 agccgtgatc gtgggcacca tcctgggctt tacactgagg ccataccgca tgtcctatcg
      901 ggaggtgaag tatttctctt ttcccggcga gctgctgatg agaatgctgc agatgctggt
      961 gctgcctctg atcatcagct ccctggtgac cggaatggcc gccctggata gcaaggcctc
     1021 cggcaagatg ggcatgagag ccgtggtgta ctatatgacc acaaccatca tcgccgtggt
     1081 catcggcatc atcatcgtga tcatcatcca ccccggcaag ggcaccaagg agaatatgca
     1141 cagggagggc aagatcgtgc gcgtgacagc agcagacgcc ttcctggatc tgatcaggaa
     1201 catgtttccc cctaatctgg tggaggcctg cttcaagcag tttaagacca actacgagaa
     1261 gcggagcttc aaggtgccta tccaggccaa tgagacactg gtgggcgccg tgatcaacaa
     1321 cgtgagcgag gccatggaga ccctgacaag gatcaccgag gagctggtgc cagtgccagg
     1381 atctgtgaac ggcgtgaatg ccctgggcct ggtggtgttc agcatgtgct tcggctttgt
     1441 gatcggcaac atgaaggagc agggccaggc cctgagggag ttctttgaca gcctgaatga
     1501 ggccatcatg cgcctggtgg ccgtgatcat gtggtatgcc cccgtgggca tcctgtttct
     1561 gatcgccggc aagatcgtgg agatggagga tatgggcgtg atcggcggcc agctggccat
     1621 gtacaccgtg acagtgatcg tgggcctgct gatccacgcc gtgatcgtgc tgcctctgct
     1681 gtatttcctg gtgacccgga agaacccatg ggtgtttatc ggaggactgc tgcaggccct
     1741 gatcaccgcc ctgggcacat ctagctcctc tgccaccctg cctatcacat tcaagtgcct
     1801 ggaggagaac aatggcgtgg acaagcgggt gaccagattt gtgctgccag tgggcgccac
     1861 catcaacatg gatggaacag ccctgtacga ggccctggca gcaatcttca tcgcccaggt
     1921 gaacaatttc gagctgaatt ttggccagat catcacaatc tctatcaccg caacagcagc
     1981 aagcatcgga gcagcaggca tcccacaggc cggcctggtg accatggtca tcgtgctgac
     2041 cagcgtggga ctgccaaccg acgatatcac actgatcatc gccgtggact ggtttctgga
     2101 taggctgcgc acaaccacaa acgtgctggg cgacagcctg ggagcaggaa tcgtggagca
     2161 cctgtcccgg cacgagctga agaatagaga tgtggagatg ggcaacagcg tgatcgagga
     2221 gaatgagatg aagaagccct atcagctgat cgcccaggac aacgagaccg agaagcctat
     2281 cgattccgag acaaagatgg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2341 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2401 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2461 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2521 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2581 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2641 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2701 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2761 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2821 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2881 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2941 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3001 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3061 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3121 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3181 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3241 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3301 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3361 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3421 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3481 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3541 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3601 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3661 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3721 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3781 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3841 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3901 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3961 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4021 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4081 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4141 gttcctggcc ttttgctggc cttttgctca catgtt
//
