LOCUS       sequence-437150-        3187 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 3187)
  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 3187)
  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: pENTR-SLC66A2_STOP
FEATURES             Location/Qualifiers
     source          1..3187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(823..922)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(979..995)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(979..995)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(992..1014)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1167..1755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1247..1266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1891..2706)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1891..2706)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2614..2633
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2698..2717
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2827..2843
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2827..2843
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      2844..2890
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3004..3026
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3018..3035
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3019..3035
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3088..3187
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ccaccatgga ggccgaaggc ctggactggc tgttggtgcc cttacatcag ctcgtgagtt
       61 ggggtgccgc tgcagctatg gtgttcggag gcgtggtacc gtatgtcccg cagtacaggg
      121 atatacgtcg cactcagaat gccgacgggt tttcaacata cgtgtgtctg gtgctgctgg
      181 ttgcaaatat tttgcggatc ctgttttggt tcggcagaag gttcgagagc cctcttctgt
      241 ggcaaagcgc cataatgatc ttgacaatgc tgcttatgct taagctctgc accgaggtga
      301 gagtcgccaa tgaattaaac gcacggcgga gatccttcac tgcggcggat agcaaagacg
      361 aggaagttaa ggtagcgcca aggagatcct tcctcgattt cgacccccac cacttttggc
      421 agtggagttc attttctgat tatgtccagt gtgtactggc attcaccggc gtggcgggct
      481 atataactta cctctcaata gattctgcat tgttcgtcga gaccctcggc ttcctcgctg
      541 tgttgactga agcaatgtta ggagtgcctc agctctaccg gaatcataga caccagtcca
      601 ccgagggaat gtctattaag atggtactga tgtggacctc tggagacgcc ttcaaaaccg
      661 catacttcct cctcaaagga gctccactgc agttttccgt ctgtggcttg ttgcaggtcc
      721 tggtggacct ggcaattttg ggccaggcat acgcgttcgc aaggcaccca cagaagccgg
      781 caccgcacgc cgtacaccct acaggcacta aagcgttgta agacccagct ttcttgtaca
      841 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
      901 gtcaaaataa aatcattatt tgaggctagg tggaggctca gtgatgataa gtctgcgatg
      961 gtggatgcat gtgtcatggt catagctgtt tcctgtgtga aattgttatc cgctcagagg
     1021 gcacaatcct attccgcgct atccgacaat ctccaagaca ttaggtggag ttcagttcgg
     1081 cgtatggcat atgtcgctgg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1141 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1201 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1261 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1321 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1381 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1441 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1501 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1561 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1621 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1681 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1741 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tctattcaac
     1801 aaagccgccg tcccgtcaag tcagcgtaaa tgggtagggg gcttcaaatc gtccgctctg
     1861 ccagtgttac aaccaattaa caaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     1921 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1981 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     2041 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     2101 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
     2161 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     2221 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     2281 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     2341 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     2401 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     2461 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     2521 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     2581 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     2641 atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg
     2701 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     2761 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttcccc
     2821 cgccgctcta gaactagtgg atccaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2881 ttcgttttat ctgttgtttg tcgcattata cgagacgtcc aggttgggat acctgaaaca
     2941 aaacccatcg tacggccaag gaagtctcca ataactgtga tccaccacaa gcgccagggt
     3001 tttcccagtc acgacgttgt aaaacgacgg ccagtcatgc ataatccgca cgcatctgga
     3061 ataaggaagt gccattccgc ctgacctcaa ataatgattt tattttgact gatagtgacc
     3121 tgttcgttgc aacagattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
     3181 gcaggct
//
