LOCUS       sequence-479341-        4176 bp    DNA     circular SYN 08-AUG-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 4176)
  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 4176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 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-SLC2A10_STOP
FEATURES             Location/Qualifiers
     source          1..4176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(681..767)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     816..838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     830..847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..962
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2595..2694)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2711..2730)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2712..2730)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2735..2751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2735..2751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2853..2872)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2864..3673
                     /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(2931..2950)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3820..4176,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
      541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
      661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
      781 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
      841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      901 gttgcaacac attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg
      961 ctccaccatg ggacactccc cacctgtgct gccactgtgc gcatccgtgt ctctgctggg
     1021 aggactgacc ttcggctacg agctggccgt gatctctggc gccctgctgc cactgcagct
     1081 ggactttggc ctgagctgcc tggagcagga gttcctggtg ggatccctgc tgctgggcgc
     1141 cctgctggcc tctctggtgg gcggctttct gatcgattgt tatggcagga agcaggccat
     1201 cctgggatcc aacctggtgc tgctggcagg atctctgacc ctgggactgg caggaagcct
     1261 ggcatggctg gtgctgggcc gcgccgtggt gggcttcgcc atcagcctga gctccatggc
     1321 ctgctgtatc tacgtgtccg agctggtggg acctaggcag agaggcgtgc tggtgtccct
     1381 gtatgaggcc ggcatcaccg tgggcatcct gctgtcttac gccctgaatt atgcactggc
     1441 aggaacccca tggggatgga ggcacatgtt tggatgggca acagcaccag ccgtgctgca
     1501 gagcctgtcc ctgctgttcc tgcctgcagg aacagacgag accgcaacac acaaggatct
     1561 gatccctctg cagggaggag aggcaccaaa gctgggacct ggcaggccac gctactcctt
     1621 tctggacctg ttccgggcca gagataacat gaggggaagg accacagtgg gactgggact
     1681 ggtgctgttt cagcagctga ccggccagcc caacgtgctg tgctatgcct ccacaatctt
     1741 ttctagcgtg ggattccacg gaggatcctc tgccgtgctg gcatctgtgg gactgggagc
     1801 agtgaaggtg gcagccaccc tgacagcaat gggactggtg gacagagcag gccggagagc
     1861 cctgctgctg gcaggatgcg ccctgatggc cctgtctgtg agcggaatcg gactggtgag
     1921 cttcgccgtg cccatggata gcggaccttc ctgtctggca gtgccaaacg caaccggaca
     1981 gacaggcctg cctggcgact ccggcctgct gcaggatagc tccctgccac ccatcccaag
     2041 gaccaatgag gaccagaggg agccaatcct gagcaccgcc aagaagacaa agccacaccc
     2101 caggagcggc gatccctccg cccctccacg cctggccctg tctagcgccc tgccaggacc
     2161 acctctgcca gcaaggggac acgcactgct gagatggacc gccctgctgt gcctgatggt
     2221 gttcgtgagc gccttcagct tcggcttcgg ccctgtgaca tggctggtgc tgtctgagat
     2281 ctacccagtg gagatccggg gcagagcctt tgccttctgt aacagcttta attgggccgc
     2341 caacctgttt atctccctgt ctttcctgga cctgatcggc accatcggcc tgagctggac
     2401 attcctgctg tacggactga ccgccgtgct gggactgggc tttatctatc tgttcgtgcc
     2461 cgagacaaag ggccagagcc tggccgagat cgatcagcag ttccagaaga ggcgctttac
     2521 cctgtccttc ggccaccggc agaattctac aggcatccct tatagcagaa tcgagatctc
     2581 cgccgcctct taagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2641 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2701 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2761 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2821 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2881 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2941 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     3001 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3061 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3121 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     3181 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3241 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3301 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3361 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3421 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3481 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3541 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3601 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3661 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3721 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3781 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3841 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3901 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3961 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4021 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4081 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4141 ggggggttcg tgcacacagc ccagcttgga gcgaac
//
