LOCUS       sequence-338286-        4545 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 4545)
  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 4545)
  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-SLC5A1_STOP
FEATURES             Location/Qualifiers
     source          1..4545
                     /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(2102..2201)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2218..2237)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2219..2237)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2242..2258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2242..2258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2360..2379)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2371..3180
                     /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(2438..2457)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3327..3915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3816..3835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4069..4086
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4245..4272)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4364..4450)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4499..4521
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4513..4530
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4514..4530
                     /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 ccaccatgga cagctccacc
      121 tggtctccta agaccacagc cgtgacccgc ccagtggaga cacacgagct gatccggaac
      181 gccgccgata tcagcatcat cgtgatctac ttcgtggtgg tcatggcagt gggactgtgg
      241 gccatgtttt ctaccaatag aggcacagtg ggcggcttct ttctggcagg caggagcatg
      301 gtgtggtggc caatcggagc aagcctgttc gcctccaaca tcggatctgg acactttgtg
      361 ggactggcag gaaccggagc agcatccggc atcgccatcg gcggcttcga gtggaatgca
      421 ctggtgctgg tggtggtgct gggatggctg tttgtgccaa tctacatcaa ggccggcgtg
      481 gtgaccatgc ccgagtatct gaggaagcgg ttcggaggac agaggatcca ggtgtacctg
      541 agcctgctgt ccctgctgct gtatatcttt acaaagatct ctgccgacat cttcagcggc
      601 gccatcttta tcaacctggc cctgggcctg aatctgtacc tggccatctt cctgctgctg
      661 gccatcaccg ccctgtacac catcacaggc ggcctggccg ccgtgatcta taccgacaca
      721 ctgcagaccg tgatcatgct ggtgggctcc ctgatcctga caggcttcgc ctttcacgaa
      781 gtgggcggct acgatgcctt catggagaag tatatgaagg ccatccccac aatcgtgtcc
      841 gacggcaaca ccacatttca ggagaagtgc tacaccccta gagccgattc tttccacatc
      901 tttagggacc cactgacagg cgatctgcca tggcccggct tcatctttgg catgtctatc
      961 ctgaccctgt ggtattggtg cacagatcag gtcatcgtgc agagatgtct gtccgccaag
     1021 aatatgtctc acgtgaaggg cggctgcatc ctgtgcggct acctgaagct gatgccaatg
     1081 ttcatcatgg tcatgcccgg catgatcagc aggatcctgt ataccgagaa gatcgcctgc
     1141 gtggtgcctt ccgagtgcga gaagtactgt ggcaccaaag tgggctgtac aaacatcgcc
     1201 tatcccaccc tggtggtgga gctgatgcct aatggcctgc gcggcctgat gctgagcgtg
     1261 atgctggcct ccctgatgtc tagcctgaca tccatcttca actccgcctc taccctgttt
     1321 acaatggaca tctacgccaa ggtgcggaag agagccagcg agaaggagct gatgatcgcc
     1381 ggccggctgt tcatcctggt gctgatcggc atctccatcg cctgggtgcc tatcgtgcag
     1441 tctgcccaga gcggccagct gtttgattac atccagtcca tcacctctta tctgggacca
     1501 cctatcgcag ccgtgttcct gctggccatc ttttggaaga gggtgaacga gccaggagca
     1561 ttctggggac tgatcctggg actgctgatc ggcatcagcc ggatgatcac cgagtttgcc
     1621 tatggcacag gctcttgcat ggagcccagc aattgcccta caatcatctg tggcgtgcac
     1681 tacctgtatt tcgccatcat cctgttcgcc atctccttta tcaccatcgt ggtcatctct
     1741 ctgctgacaa agcctatccc agacgtgcac ctgtacaggc tgtgctggag cctgcgcaac
     1801 tccaaggagg agcggatcga cctggatgcc gaggaggaga atatccagga gggccccaag
     1861 gagaccatcg agatcgagac acaggtgcct gagaagaaga agggcatctt ccggagagcc
     1921 tatgacctgt tttgcggact ggagcagcac ggagcaccaa agatgaccga ggaggaggag
     1981 aaggccatga agatgaagat gaccgataca agcgagaagc ccctgtggag aaccgtgctg
     2041 aacgtgaatg gcatcatcct ggtgacagtg gccgtgttct gtcacgccta ctttgcctaa
     2101 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     2161 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     2221 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     2281 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     2341 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     2401 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     2461 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     2521 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     2581 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     2641 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     2701 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     2761 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     2821 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     2881 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     2941 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     3001 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     3061 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     3121 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     3181 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     3241 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     3301 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3361 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3421 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3481 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3541 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3601 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3661 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3721 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3781 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3841 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3901 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3961 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4021 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4081 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4141 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     4201 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     4261 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     4321 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     4381 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     4441 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     4501 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
