LOCUS       sequence-274433-        5460 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 5460)
  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 5460)
  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_SLC26A8
FEATURES             Location/Qualifiers
     source          1..5460
                     /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(3585..3684)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3701..3720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3702..3720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3725..3741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3725..3741)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3843..3862)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3854..4663
                     /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(3921..3940)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4810..5398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5299..5318
                     /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 accatggcac agctggagcg gtctgccatc agcggcttta gctccaagtc
      721 ccggagaaac tctttcgcct acgatgtgaa gagggaggtg tataatgagg agaccttcca
      781 gcaggagcac aagcgcaagg cctctagctc cggcaacatg aatatcaaca tcaccacatt
      841 tcgccaccac gtgcagtgcc ggtgtagctg gcacagattc ctgagatgcg tgctgacaat
      901 cttcccattt ctggagtgga tgtgcatgta caggctgaag gattggctgc tgggcgacct
      961 gctggcagga atcagcgtgg gactggtgca ggtgccacag ggactgaccc tgtccctgct
     1021 ggcaaggcag ctgatcccac ctctgaacat cgcctacgcc gccttttgct ctagcgtgat
     1081 ctatgtgatc ttcggcagct gtcaccagat gtccatcggc tctttctttc tggtgagcgc
     1141 cctgctgatc aatgtgctga aggtgtcccc ttttaacaat ggccagctgg tcatgggctc
     1201 cttcgtgaag aacgagttta gcgccccatc ctacctgatg ggctataata agagcctgtc
     1261 cgtggtggcc accacaacct ttctgacagg catcatccag ctgatcatgg gcgtgctggg
     1321 actgggcttc atcgcaacct acctgcccga gtctgccatg agcgcctatc tggcagcagt
     1381 ggccctgcac atcatgctga gccagctgac attcatcttt ggcatcatga tctctttcca
     1441 cgccggccct atcagcttct tttacgacat catcaattat tgcgtggccc tgccaaaggc
     1501 caactctacc agcatcctgg tgtttctgac agtggtggtg gccctgagaa tcaataagtg
     1561 tatcaggatc tccttcaacc agtaccccat cgagttccct atggagctgt ttctgatcat
     1621 cggcttcacc gtgatcgcca acaagatcag catggccaca gagacctccc agaccctgat
     1681 cgatatgatc ccctattctt ttctgctgcc agtgacaccc gacttcagcc tgctgcctaa
     1741 gatcatcctg caggccttct ccctgtctct ggtgtcctct ttcctgctga tctttctggg
     1801 caagaagatc gccagcctgc acaactactc cgtgaattct aaccaggatc tgatcgccat
     1861 cggcctgtgc aatgtggtga gctccttctt tagatcctgc gtgttcaccg gcgccatcgc
     1921 caggacaatc atccaggaca agagcggagg ccgccagcag ttcgcatccc tggtgggcgc
     1981 cggcgtgatg ctgctgctga tggtgaagat gggccacttc ttttatacac tgcctaatgc
     2041 cgtgctggcc ggcatcatcc tgagcaacgt gatcccatac ctggagacca tcagcaatct
     2101 gccctccctg tggaggcagg atcagtatga ctgcgccctg tggatgatga ccttttctag
     2161 ctccatcttc ctgggcctgg atatcggcct gatcatcagc gtggtgtccg ccttctttat
     2221 cacaaccgtg cgcagccaca gggcaaagat cctgctgctg ggacagatcc caaataccaa
     2281 catctaccgc tccatcaacg actatcggga gatcatcaca atccccggcg tgaagatctt
     2341 tcagtgctgt tctagcatca ccttcgtgaa cgtgtactat ctgaagcaca agctgctgaa
     2401 ggaggtggat atggtgaagg tgcctctgaa ggaggaggag atcttcagcc tgtttaattc
     2461 ctctgacaca aacctgcagg gcggcaagat ctgcaggtgt ttttgcaact gtgacgatct
     2521 ggagccactg cccagaatcc tgtacaccga gaggttcgag aacaagctgg atccagaggc
     2581 cagctccatc aatctgatcc actgctctca ctttgagagc atgaatacct cccagacagc
     2641 ctctgaggac caggtgccct acacagtgtc tagcgtgagc cagaagaacc agggccagca
     2701 gtatgaggaa gtggaggaag tgtggctgcc taacaattcc tctcgcaata gctcccctgg
     2761 cctgccagat gtggcagagt ctcagggcag gcgcagcctg atcccttact ctgacgccag
     2821 cctgctgcca tccgtgcaca ccatcatcct ggatttctcc atggtgcact atgtggactc
     2881 taggggactg gtggtgctga ggcagatctg caacgccttt cagaatgcca acatcctgat
     2941 cctgatcgcc ggctgtcact ctagcatcgt gagagccttc gagaggaatg atttctttga
     3001 cgccggcatc acaaagaccc agctgtttct gtccgtgcac gatgccgtgc tgttcgccct
     3061 gtctcggaaa gtgatcggct cctctgagct gagcatcgac gagtccgaga ccgtgatccg
     3121 ggagacctac tctgagacag ataagaatga caacagcaga tataagatga gctcctcttt
     3181 tctgggctcc cagaagaacg tgtctcccgg cttcatcaag atccagcagc ctgtggagga
     3241 ggagtccgag ctggacctgg agctggagtc tgagcaggag gcaggactgg gactggacct
     3301 ggatctggac agagagctgg agccagagat ggagcccaag gccgagacag agaccaagac
     3361 acagaccgag atggagcctc agcctgagac agagcccgag atggagccca accctaagag
     3421 caggccaagg gcacacacct tccctcagca gcggtactgg cctatgtatc acccatccat
     3481 ggcctctacc cagtcccaga cacagaccag aacatggtct gtggagcgga gaaggcaccc
     3541 aatggatagc tactcccccg agggcaattc caacgaggac gtggacccag ctttcttgta
     3601 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     3661 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     3721 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     3781 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     3841 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     3901 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3961 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     4021 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     4081 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     4141 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     4201 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     4261 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     4321 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     4381 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     4441 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     4501 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     4561 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     4621 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     4681 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     4741 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
