LOCUS       sequence-356919-        4275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4275)
  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 4275)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1832..1931)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1948..1967)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1949..1967)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1972..1988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1972..1988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2090..2109)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2101..2910
                     /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(2168..2187)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3057..3645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3546..3565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3799..3816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3975..4002)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4094..4180)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4229..4251
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4243..4260
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4244..4260
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc cagcaccgag
      121 ggcgccaaca atatgccaaa gcaggtggag gtgaggatgc acgacagcca cctgggatcc
      181 gaggagccaa agcacaggca cctgggactg aggctgtgcg ataagctggg caagaatctg
      241 ctgctgaccc tgacagtgtt cggcgtgatc ctgggagccg tgtgcggagg cctgctgagg
      301 ctggcctctc ccatccaccc tgacgtggtc atgctgatcg cctttcccgg cgatatcctg
      361 atgcgcatgc tgaagatgct gatcctgcct ctgatcatca gctccctgat cacaggcctg
      421 agcggactgg acgcaaaggc atccggccgg ctgggcacca gagctatggt gtactatatg
      481 agcaccacaa tcatcgcagc cgtgctgggc gtgatcctgg tgctggccat ccacccaggc
      541 aaccccaagc tgaagaagca gctgggcccc ggcaagaaga atgatgaggt gtctagcctg
      601 gacgccttcc tggatctgat ccgcaacctg tttcctgaga atctggtgca ggcctgtttc
      661 cagcagatcc agaccgtgac aaagaaggtg ctggtggcac cacctccaga cgaggaggcc
      721 aacgccacat ctgccgtggt gagcctgctg aatgagaccg tgacagaggt gcctgaggag
      781 accaagatgg tcatcaagaa gggcctggag ttcaaggatg gcatgaatgt gctgggcctg
      841 atcggcttct ttatcgcctt tggcatcgcc atgggcaaga tgggcgacca ggccaagctg
      901 atggtggatt tctttaacat cctgaatgag atcgtgatga agctggtcat catgatcatg
      961 tggtactccc cactgggcat cgcctgcctg atctgtggca agatcatcgc catcaaggac
     1021 ctggaggtgg tggcaaggca gctgggcatg tacatggtga cagtgatcat cggcctgatc
     1081 atccacggcg gcatcttcct gcctctgatc tattttgtgg tgaccagaaa gaacccattc
     1141 tctttctttg ccggcatctt tcaggcatgg atcaccgccc tgggaacagc atcctctgcc
     1201 ggcaccctgc cagtgacatt ccggtgcctg gaggagaacc tgggcatcga caagagggtg
     1261 acacgctttg tgctgcccgt gggcgccacc atcaatatgg atggaacagc cctgtacgag
     1321 gcagtggcag caatcttcat cgcccagatg aacggagtgg tgctggacgg aggacagatc
     1381 gtgaccgtgt ctctgaccgc cacactggca agcgtgggag cagcctccat cccttctgcc
     1441 ggcctggtga ccatgctgct gatcctgaca gccgtgggcc tgccaaccga ggacatcagc
     1501 ctgctggtgg cagtggactg gctgctggat cggatgagaa catccgtgaa tgtggtgggc
     1561 gattcttttg gcgccggcat cgtgtatcac ctgagcaagt ccgagctgga caccatcgat
     1621 agccagcacc gggtgcacga ggatatcgag atgaccaaga cacagtccat ctacgacgat
     1681 atgaagaacc acagagagtc taacagcaat cagtgcgtgt atgccgccca caactccgtg
     1741 atcgtggacg agtgtaaggt gaccctggcc gccaatggca agtccgccga ttgctctgtg
     1801 gaggaggagc cttggaagag ggagaagtaa gacccagctt tcttgtacaa agttggcatt
     1861 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1921 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     1981 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2041 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2101 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2161 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2221 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2281 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2341 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2401 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2461 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2521 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2581 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2641 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2701 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2761 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2821 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2881 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     2941 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3001 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3841 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3901 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     3961 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     4021 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     4081 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     4141 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     4201 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     4261 cttaagctcg ggccc
//
