LOCUS       sequence-479347-        4926 bp    DNA     circular SYN 25-JUL-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 4926)
  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 4926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 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-SLC26A9_STOP
FEATURES             Location/Qualifiers
     source          1..4926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(919..1728)
                     /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     1642..1661
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1720..1739
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1841..1857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1841..1857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1862..1881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1862..1880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1898..1997
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4380..4479)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4495..4512)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4495..4511)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4504..4526)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      4575..4661
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4753..4780
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga
      841 ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa
      901 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      961 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1021 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1081 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     1141 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     1201 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     1261 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     1321 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1381 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt
     1441 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1501 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1561 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     1621 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1681 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt
     1741 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     1801 aatgtaacat cagagatttt gagacacggg ccagagctgc caggaaacag ctatgaccat
     1861 gtaatacgac tcactatagg ggatatcagc tggatggcaa ataatgattt tattttgact
     1921 gatagtgacc tgttcgttgc aacaaattga taagcaatgc tttcttataa tgccaacttt
     1981 gtacaagaaa gctgggtctt acagggctgt cagggtctcg gcgtgaaaca tgctgccgaa
     2041 catctcctgc tccagatccc aataagagcg gatatcctcc tcggagtcgt acaggctcag
     2101 ctctgcgtcg ccaggtgctc cctggaaatt gtgtcctggg gtcacatccc ttgcgtttgc
     2161 ctgtgcaaac agcacggcgt cgtggatgga ggggaacacg tgcttacact ccaggctgcc
     2221 atcctcaaac acgccgccgt gagagatgtc gttatacacc tgggcgtgga tattcaccag
     2281 gaacaccttc acgccgatct tgccgtaggt ggagctcagc ttggccaggg ccttgatgcc
     2341 catcagatcc acgaagctca cgccagacat gtccaggatc agtgtgtgaa aggtcacgaa
     2401 tggaggcacg ctggccagca tatcagatgg ctctcctggt gcctcggcgg atgctggtgg
     2461 ctcgctctgt gcaggagagg agctgtctgg gctgaatgtg atgtaagaca cggatgtgcc
     2521 attggctggg gtctggttat tgttaggatc ggtagggggg gcgttctcaa agtcctgctg
     2581 cagctcctgc agggacactg tcttggtctt catgaacagg ctgcgcctct gctgggtagg
     2641 tctcattctc cgcttctcct gcttcttcag gtacttctgc ttggccagca gcaccttctg
     2701 gggatccatg cctgtcttgg cgatcacttt ctgccgaaag atctcgctat tggcgaaata
     2761 caggggagag cagtaggtga tgatcttgat gccctggatg tcctgggctc tgttatatgt
     2821 ctttggattc acgtagatat cggtgtccat gacctgggcc agggcatagc cgttccgaaa
     2881 ctgtgtctgg aacaccacca ccagcacgct aaaggccact cccactgcca ctccgtatgg
     2941 cagagacaga aagaaagagg acaggaagga caccacccag atacagcaat ccagcttgga
     3001 ctttctccac agatagtatg ggtcggtcag ctgcttcagg gagttcttca gattcacggc
     3061 gatcagggcg cccagcacgc tcttaggcag tggatacagg tagatgccca gcaccagcat
     3121 tgtgatcatg accaccaggc tcacgcacag ggatgccacc tggctcttgc ctcctgctcc
     3181 atccactgcc agggtcacag acagggcaca gcagatcacg tggatcttaa agaaagagcc
     3241 aaagaaattg gagcagccca gggcgatcat ctcctgattg gaatccacgt catagccgtg
     3301 cttgttggcc agggtccggc ccatggccag attgatcacg tagctcacga tggccagaga
     3361 aaaggctgtg ccgatcatgt ccttccactg agacaccact ggggacactg gggtagggaa
     3421 tcccctctgg atctctccca cgatctgcat gtgatacttc ttaggcatct tacagcctcc
     3481 gctgattgct gttgccacca ccaccacgat catctcggtg gggattggga atctgatctt
     3541 gtgcatgtac cgggcgttca gctccttcac cagcaccaga aaggcgccgg agatcagggc
     3601 gaagatcagg ctggcgatgt ttgtgtgggg cagattcttg cagatgtcga taaaggtgaa
     3661 cacgatagag cctgggcctg tataggaagg gatggtcagg ccaaagatgt acttcagcac
     3721 ggagatcagg atctgcagtc ctgctgcggt cataaagccc ctgatgaagc tctcagacag
     3781 atagatggcc acaaagccga actgcatgaa gcccaggccc atctggatga tggctgtcag
     3841 acatgccagg gtggcgctca cgtgcagcct ctctgcctcc attgctgctg tgtccacgta
     3901 agactcattg gtggcattgt taaacacctg gaacttggac tcaggggcca gctgcaggca
     3961 gatgttgccc accaggatgg agatcacggc aaatgtgcct ggcaccatct ggtgcactcc
     4021 tcccagaaag aaataggtca gcaggggaaa gaagctagag tacaggccgt tcacggctgg
     4081 cagattggcc agcagggcga aggccattcc ctgtggcacc tggatggatc ctccgctcag
     4141 tcctcccagc agatcaggga tgatatagtc cttgatcttg tactttggca gccaggacag
     4201 cactggcagc agtccaaaca ccacggcctt gatcttggcg gagctgcagc ggaaggcgtt
     4261 cctcagcttc tcgcccacag ggtatgtgcg atccttcttc tcaaactcat cgtcgaacag
     4321 ggtcaggcta taggcggccc tgtccaccac gtatctaggc cggggctgag acatggtgga
     4381 gcctgctttt ttgtacaaag ttggcattat aaaaaagcat tgctcatcaa tgtgttgcaa
     4441 cgaacaggtc actatcagtc aaaataaaat cattatttgg ggcccgagct taagactggc
     4501 cgtcgtttta caacgtcgtg actgggaaaa catccatgct agcgttaacg cgagagtagg
     4561 gaactgccag gcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta
     4621 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     4681 acgttgtgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     4741 atcaaactaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttc
     4801 ctggctagcg gtacgcgtat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4921 tgggcg
//
