LOCUS       sequence-259577-        4095 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 4095)
  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 4095)
  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_SLC16A10
FEATURES             Location/Qualifiers
     source          1..4095
                     /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"
     RBS             1654..1662
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(2220..2319)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2336..2355)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2337..2355)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2360..2376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2360..2376)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2478..2497)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2489..3298
                     /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(2556..2575)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3445..4033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3934..3953
                     /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 accatggtgc tgtcccagga ggagccagat agcgccaggg gcacctccga
      721 ggcccagcct ctgggccctg ccccaacagg agcagcacca cctccaggac caggaccttc
      781 cgactctcct gaggcagccg tggagaaggt ggaggtggag ctggcaggac cagcaaccgc
      841 agagccccac gagccccctg agccacccga gggcggctgg ggctggctgg tcatgctggc
      901 cgccatgtgg tgcaacggct ccgtgttcgg catccagaat gcctgtggcg tgctgtttgt
      961 gagcatgctg gagaccttcg gctccaagga cgatgacaag atggtgttta agacagcctg
     1021 ggtgggcagc ctgtccatgg gcatgatctt cttttgctgt cccatcgtga gcgtgtttac
     1081 cgatctgttc ggctgcagga agacagcagt ggtgggagca gcagtgggct ttgtgggcct
     1141 gatgagctcc tctttcgtga gctccatcga gccactgtac ctgacatatg gcatcatctt
     1201 tgcctgcggc tgttccttcg cctaccagcc ctctctggtc atcctgggcc actacttcaa
     1261 gaagcggctg ggactggtga acggaatcgt gaccgcaggc tctagcgtgt tcacaatcct
     1321 gctgccactg ctgctgaggg tgctgatcga ttccgtgggc ctgttttaca ccctgcgggt
     1381 gctgtgcatc tttatgttcg tgctgtttct ggccggcttc acatatcggc ccctggccac
     1441 ctctacaaag gacaaggagt ccggcggctc tggctcctct ctgttttcta gaaagaagtt
     1501 cagccctcca aagaagatct ttaatttcgc catcttcaag gtgaccgcat acgccgtgtg
     1561 ggcagtggga atccctctgg ccctgtttgg ctacttcgtg ccatatgtgc acctgatgaa
     1621 gcacgtgaac gagcggttcc aggatgagaa gaataaggag gtggtgctga tgtgcatcgg
     1681 agtgacatct ggagtgggcc ggctgctgtt tggcagaatc gccgactacg tgcctggcgt
     1741 gaagaaggtg tatctgcagg tgctgagctt ctttttcatc ggcctgatgt ctatgatgat
     1801 cccactgtgc agcatcttcg gcgccctgat cgccgtgtgc ctgatcatgg gcctgtttga
     1861 tggctgtttc atctccatca tggcaccaat cgcctttgag ctggtgggag cacaggacgt
     1921 gagccaggcc atcggctttc tgctgggctt catgtccatc cctatgaccg tgggaccacc
     1981 tatcgcagga ctgctgaggg ataagctggg cagctacgac gtggcctttt atctggcagg
     2041 agtgccacca ctgatcggag gagccgtgct gtgcttcatc ccatggatcc actccaagaa
     2101 gcagcgcgag atctctaaga ccacaggcaa ggagaagatg gagaagatgc tggagaacca
     2161 gaatagcctg ctgagctcct ctagcggcat gttcaagaag gagtctgaca gcatcatcga
     2221 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2281 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2341 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2401 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2461 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2521 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2581 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2641 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2701 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2761 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2821 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2881 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2941 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3001 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3061 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3121 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3181 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3241 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3301 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3361 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3421 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3481 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3541 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3601 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3661 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3721 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3781 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3841 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3901 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3961 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4021 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4081 ttttgctcac atgtt
//
