LOCUS       sequence-253155-        4122 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 4122)
  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 4122)
  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_SLC1A1
FEATURES             Location/Qualifiers
     source          1..4122
                     /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(2247..2346)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2363..2382)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2364..2382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2387..2403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2387..2403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2505..2524)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2516..3325
                     /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(2583..2602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3472..4060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3961..3980
                     /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 accatgggca agcctgccag gaagggctgc gagtggaagc ggttcctgaa
      721 gaacaattgg gtgctgctgt ctaccgtggc agcagtggtg ctgggaatca ccacaggcgt
      781 gctggtgagg gagcactcca acctgtctac actggagaag ttctactttg ccttccctgg
      841 cgagatcctg atgcgcatgc tgaagctgat catcctgcca ctgatcatca gctccatgat
      901 caccggagtg gccgccctgg atagcaacgt gagcggcaag atcggcctga gggccgtggt
      961 gtactatttc tgcaccacac tgatcgccgt gatcctgggc atcgtgctgg tggtgagcat
     1021 caagcccggc gtgacccaga aagtgggaga gatcgcaagg accggatcca cacctgaggt
     1081 gtctacagtg gacgccatgc tggatctgat cagaaacatg tttcccgaga atctggtgca
     1141 ggcctgtttc cagcagtaca agaccaagcg ggaggaggtg aagccccctt ctgatcctga
     1201 gatgaacatg accgaggaga gcttcacagc cgtgatgacc acagccatct ccaagaacaa
     1261 gacaaaggag tacaagatcg tgggcatgta tagcgacggc atcaatgtgc tgggcctgat
     1321 cgtgttttgc ctggtgttcg gcctggtcat cggcaagatg ggcgagaagg gccagatcct
     1381 ggtggacttc tttaatgccc tgtccgatgc caccatgaag atcgtgcaga tcatcatgtg
     1441 ctatatgcca ctgggcatcc tgtttctgat cgccggcaag atcatcgagg tggaggattg
     1501 ggagatcttc cggaagctgg gcctgtacat ggccaccgtg ctgacaggcc tggccatcca
     1561 ctccatcgtg atcctgccac tgatctactt catcgtggtg cggaagaacc cctttagatt
     1621 cgccatggga atggcacagg ccctgctgac cgccctgatg atctctagct cctctgccac
     1681 cctgcctgtg accttccggt gtgccgagga gaacaatcag gtggacaaga ggatcacacg
     1741 cttcgtgctg ccagtgggcg ccaccatcaa catggatgga acagccctgt acgaggcagt
     1801 ggcagccgtg ttcatcgcac agctgaatga cctggatctg ggcatcggcc agatcatcac
     1861 catctccatc accgccacat ctgccagcat cggagcagca ggagtgccac aggcaggact
     1921 ggtgaccatg gtcatcgtgc tgtctgccgt gggactgcca gcagaggacg tgacactgat
     1981 catcgccgtg gactggctgc tggatcggtt tagaaccatg gtgaatgtgc tgggcgacgc
     2041 cttcggcaca ggcatcgtgg agaagctgtc taagaaggag ctggagcaga tggatgtgag
     2101 ctccgaggtg aacatcgtga atccctttgc cctggagagc accatcctgg acaacgagga
     2161 cagcgataca aagaagtcct atgtgaatgg cggcttcgcc gtggacaagt ctgataccat
     2221 cagctttacc cagacaagcc agttcgaccc agctttcttg tacaaagttg gcattataag
     2281 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2341 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2401 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaataat
     2461 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2521 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2581 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2641 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2701 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2761 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2821 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2881 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2941 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3001 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3061 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3121 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3181 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3241 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3301 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3361 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3421 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3601 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4081 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
