LOCUS       sequence-259227-        4272 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 4272)
  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 4272)
  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_SLC1A2
FEATURES             Location/Qualifiers
     source          1..4272
                     /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(2397..2496)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2513..2532)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2514..2532)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2537..2553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2537..2553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2655..2674)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2666..3475
                     /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(2733..2752)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3622..4210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4111..4130
                     /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 accatggcca gcaccgaggg cgccaacaat atgccaaagc aggtggaggt
      721 gaggatgcac gacagccacc tgggatccga ggagccaaag cacaggcacc tgggactgag
      781 gctgtgcgat aagctgggca agaatctgct gctgaccctg acagtgttcg gcgtgatcct
      841 gggagccgtg tgcggaggcc tgctgaggct ggcctctccc atccaccctg acgtggtcat
      901 gctgatcgcc tttcccggcg atatcctgat gcgcatgctg aagatgctga tcctgcctct
      961 gatcatcagc tccctgatca caggcctgag cggactggac gcaaaggcat ccggccggct
     1021 gggcaccaga gctatggtgt actatatgag caccacaatc atcgcagccg tgctgggcgt
     1081 gatcctggtg ctggccatcc acccaggcaa ccccaagctg aagaagcagc tgggccccgg
     1141 caagaagaat gatgaggtgt ctagcctgga cgccttcctg gatctgatcc gcaacctgtt
     1201 tcctgagaat ctggtgcagg cctgtttcca gcagatccag accgtgacaa agaaggtgct
     1261 ggtggcacca cctccagacg aggaggccaa cgccacatct gccgtggtga gcctgctgaa
     1321 tgagaccgtg acagaggtgc ctgaggagac caagatggtc atcaagaagg gcctggagtt
     1381 caaggatggc atgaatgtgc tgggcctgat cggcttcttt atcgcctttg gcatcgccat
     1441 gggcaagatg ggcgaccagg ccaagctgat ggtggatttc tttaacatcc tgaatgagat
     1501 cgtgatgaag ctggtcatca tgatcatgtg gtactcccca ctgggcatcg cctgcctgat
     1561 ctgtggcaag atcatcgcca tcaaggacct ggaggtggtg gcaaggcagc tgggcatgta
     1621 catggtgaca gtgatcatcg gcctgatcat ccacggcggc atcttcctgc ctctgatcta
     1681 ttttgtggtg accagaaaga acccattctc tttctttgcc ggcatctttc aggcatggat
     1741 caccgccctg ggaacagcat cctctgccgg caccctgcca gtgacattcc ggtgcctgga
     1801 ggagaacctg ggcatcgaca agagggtgac acgctttgtg ctgcccgtgg gcgccaccat
     1861 caatatggat ggaacagccc tgtacgaggc agtggcagca atcttcatcg cccagatgaa
     1921 cggagtggtg ctggacggag gacagatcgt gaccgtgtct ctgaccgcca cactggcaag
     1981 cgtgggagca gcctccatcc cttctgccgg cctggtgacc atgctgctga tcctgacagc
     2041 cgtgggcctg ccaaccgagg acatcagcct gctggtggca gtggactggc tgctggatcg
     2101 gatgagaaca tccgtgaatg tggtgggcga ttcttttggc gccggcatcg tgtatcacct
     2161 gagcaagtcc gagctggaca ccatcgatag ccagcaccgg gtgcacgagg atatcgagat
     2221 gaccaagaca cagtccatct acgacgatat gaagaaccac agagagtcta acagcaatca
     2281 gtgcgtgtat gccgcccaca actccgtgat cgtggacgag tgtaaggtga ccctggccgc
     2341 caatggcaag tccgccgatt gctctgtgga ggaggagcct tggaagaggg agaaggaccc
     2401 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2461 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2521 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2581 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2641 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2701 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2761 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2821 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2881 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2941 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     3001 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3061 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3121 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3181 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3241 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3301 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3361 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3421 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3481 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3541 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3601 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3661 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3721 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3781 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3841 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3901 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3961 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4021 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4081 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4141 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4201 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4261 tgctcacatg tt
//
