LOCUS       sequence-261200-        4290 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 4290)
  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 4290)
  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_SLC5A7
FEATURES             Location/Qualifiers
     source          1..4290
                     /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(2415..2514)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2531..2550)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2532..2550)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2555..2571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2555..2571)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2673..2692)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2684..3493
                     /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(2751..2770)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3640..4228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4129..4148
                     /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 accatggcct tccacgtgga gggcctgatc gccatcatcg tgttttacct
      721 gctgatcctg ctggtgggaa tctgggcagc atggaggaca aagaacagcg gatccgccga
      781 ggagaggagc gaggcaatca tcgtgggagg aagggacatc ggactgctgg tgggcggctt
      841 caccatgaca gcaacctggg tgggaggagg ctacatcaat ggaaccgcag aggccgtgta
      901 cgtgccagga tatggactgg catgggcaca ggcacctatc ggatattctc tgagcctgat
      961 cctgggcggc ctgttctttg ccaagccaat gcggtccaag ggctacgtga ccatgctgga
     1021 tcccttccag cagatctatg gcaagagaat gggcggcctg ctgtttatcc ctgccctgat
     1081 gggcgagatg ttctgggcag cagcaatctt tagcgccctg ggagccacaa tctccgtgat
     1141 catcgacgtg gatatgcaca tctccgtgat catctctgcc ctgatcgcca cactgtacac
     1201 cctggtggga ggactgtaca gcgtggccta taccgacgtg gtgcagctgt tctgcatctt
     1261 tgtgggcctg tggatctctg tgcccttcgc cctgagccac cctgcagtgg ccgatatcgg
     1321 ctttacagcc gtgcacgcca agtaccagaa gccatggctg ggaaccgtgg acagctccga
     1381 ggtgtatagc tggctggatt ccttcctgct gctgatgctg ggcggcatcc cttggcaggc
     1441 ctactttcag cgggtgctgt ctagctcctc tgccacctat gcccaggtgc tgtctttcct
     1501 ggccgccttt ggctgcctgg tcatggccat cccagcaatc ctgatcggag caatcggagc
     1561 cagcacagac tggaaccaga ccgcctacgg cctgccagac cccaagacca cagaggaggc
     1621 cgatatgatc ctgcctatcg tgctgcagta cctgtgccca gtgtatatct ctttctttgg
     1681 actgggagcc gtgagcgccg ccgtgatgag ctccgccgac tctagcatcc tgagcgcctc
     1741 ctctatgttc gcccggaaca tctaccagct gtcctttaga cagaatgcct ctgataagga
     1801 gatcgtgtgg gtcatgcgga tcacagtgtt cgtgtttgga gcatccgcca ccgcaatggc
     1861 actgctgaca aagaccgtgt acggcctgtg gtatctgagc tccgatctgg tgtatatcgt
     1921 gatcttccca cagctgctgt gcgtgctgtt tgtgaagggc acaaatacct acggagcagt
     1981 ggcaggatac gtgagcggac tgttcctgag aatcacaggc ggcgagccat acctgtatct
     2041 gcagcccctg atcttttacc ctggctacta tccagacgat aacggcatct ataatcagaa
     2101 gttccccttt aagaccctgg ccatggtgac atccttcctg accaacatct gtatctctta
     2161 cctggccaag tatctgttcg agagcggcac actgccccct aagctggacg tgtttgatgc
     2221 cgtggtggcc aggcactccg aggagaatat ggacaagacc atcctggtga agaacgagaa
     2281 tatcaagctg gatgagctgg ccctggtgaa gcctcgccag tccatgacac tgtctagcac
     2341 attcaccaac aaggaggcct ttctggacgt ggattcctct ccagagggct ctggcaccga
     2401 ggacaatctg caggacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2461 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2521 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2581 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2641 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2701 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2761 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2821 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2881 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2941 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     3001 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3061 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3121 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3181 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3241 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3301 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3361 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3421 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3481 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3541 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3601 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3661 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3721 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3781 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3841 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3901 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3961 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4021 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4081 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4141 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4201 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4261 ggccttttgc tggccttttg ctcacatgtt
//
