LOCUS       sequence-261832-        4782 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 4782)
  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 4782)
  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_SLC26A5
FEATURES             Location/Qualifiers
     source          1..4782
                     /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(2907..3006)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3023..3042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3024..3042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3047..3063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3047..3063)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3165..3184)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3176..3985
                     /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(3243..3262)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4132..4720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4621..4640
                     /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 accatggacc acgccgagga gaatgagatc ctggccgcca cccagaggta
      721 ctatgtggag cgccccatct tcagccaccc tgtgctgcag gagaggctgc acacaaagga
      781 caaggtgccc gattccatcg ccgacaagct gaagcaggcc tttacctgca cacctaagaa
      841 gatccgcaac atcatctata tgttcctgcc aatcaccaag tggctgcccg cctacaagtt
      901 taaggagtat gtgctgggcg atctggtgtc tggcatcagc acaggcgtgc tgcagctgcc
      961 tcagggactg gccttcgcca tgctggcagc agtgccacct atcttcggac tgtacagctc
     1021 cttttatcca gtgatcatgt actgtttcct gggcaccagc aggcacatct ccatcggacc
     1081 ctttgccgtg atcagcctga tgatcggagg agtggccgtg agactggtgc ctgacgatat
     1141 cgtgatccca ggcggcgtga acgcaaccaa tggaacagag gcaagggacg cactgagagt
     1201 gaaggtggcc atgtctgtga ccctgctgag cggcatcatc cagttctgcc tgggcgtgtg
     1261 ccggttcggc tttgtggcca tctacctgac agagcccctg gtgagaggct ttaccacagc
     1321 cgccgccgtg cacgtgttca cctccatgct gaagtatctg tttggcgtga agacaaagag
     1381 gtactctggc atcttcagcg tggtgtattc caccgtggcc gtgctgcaga atgtgaagaa
     1441 cctgaacgtg tgctctctgg gcgtgggcct gatggtgttt ggcctgctgc tgggcggcaa
     1501 ggagttcaac gagcggttca aggagaagct gcctgcccca atccccctgg agttctttgc
     1561 agtggtcatg ggaaccggaa tcagcgccgg cttcaatctg aaggagtcct acaacgtgga
     1621 cgtggtgggc accctgccac tgggactgct gccacccgcc aatcctgaca caagcctgtt
     1681 ccacctggtg tatgtggatg ccatcgccat cgccatcgtg ggcttttctg tgaccatcag
     1741 catggccaag acactggcca ataagcacgg ctaccaggtg gatggcaacc aggagctgat
     1801 cgccctgggc ctgtgcaact ccatcggctc tctgttccag accttttcca tctcttgtag
     1861 cctgtccaga tctctggtgc aggagggaac cggaggcaag acacagctgg caggatgtct
     1921 ggcctccctg atgatcctgc tggtcatcct ggccacaggc ttcctgtttg agtccctgcc
     1981 acaggccgtg ctgtctgcca tcgtgatcgt gaatctgaag ggcatgttca tgcagttttc
     2041 cgacctgcct ttcttttggc ggacctctaa gatcgagctg acaatctggc tgaccacatt
     2101 cgtgtctagc ctgtttctgg gcctggatta cggcctgatc accgccgtga tcatcgccct
     2161 gctgaccgtg atctacagaa cacagagccc atcctataag gtgctgggca agctgcccga
     2221 gacagacgtg tacatcgaca tcgatgccta tgaggaggtg aaggagatcc ctggcatcaa
     2281 gatcttccag atcaatgccc caatctacta tgccaattcc gatctgtact ctaacgccct
     2341 gaagaggaag accggagtga acccagccgt gatcatggga gcaaggagaa aggccatgcg
     2401 caagtatgcc aaggaagtgg gcaacgccaa tatggcaaac gcaaccgtgg tgaaggcaga
     2461 cgcagaggtg gacggagagg atgccacaaa gcctgaggag gaggatggcg aggtgaagta
     2521 tcctccaatc gtgatcaagt ccaccttccc agaggagatg cagcggttta tgccccctgg
     2581 cgataatgtg cacaccgtga tcctggactt cacacaggtg aactttatcg attctgtggg
     2641 cgtgaagacc ctggccggca tcgtgaagga gtacggcgac gtgggcatct acgtgtatct
     2701 ggcaggatgc agcgcccagg tggtgaacga tctgacaagg aatcgcttct ttgagaaccc
     2761 agccctgtgg gagctgctgt ttcactctat ccacgacgcc gtgctgggca gccagctgcg
     2821 ggaggccctg gcagagcagg aggcaagcgc cccaccatcc caggaggatc tggagcccaa
     2881 cgccacccct gccacaccag aggccgaccc agctttcttg tacaaagttg gcattataag
     2941 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     3001 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     3061 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     3121 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     3181 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     3241 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     3301 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     3361 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     3421 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     3481 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3541 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3601 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3661 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3721 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3781 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3841 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3901 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3961 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     4021 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     4081 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4741 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
