LOCUS       sequence-260114-        4239 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 4239)
  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 4239)
  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_SLC22A6
FEATURES             Location/Qualifiers
     source          1..4239
                     /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(2364..2463)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2480..2499)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2481..2499)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2504..2520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2504..2520)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2622..2641)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2633..3442
                     /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(2700..2719)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3589..4177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4078..4097
                     /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 accatggcct tcaacgacct gctgcagcaa gtgggaggag tgggccggtt
      721 tcagcagatc caggtgaccc tggtggtgct gcctctgctg ctgatggcca gccacaacac
      781 cctgcagaat ttcacagccg ccatcccaac ccaccactgc aggccacctg cagacgcaaa
      841 cctgtccaag aatggaggac tggaagtgtg gctgcctagg gatagacagg gccagccaga
      901 gagctgtctg agattcacat ccccacagtg gggcctgccc tttctgaacg gaaccgaggc
      961 aaatggaacc ggagcaacag agccatgcac agacggctgg atctacgata attctacctt
     1021 ccctagcacc atcgtgacag agtgggacct ggtgtgcagc cacagggccc tgaggcagct
     1081 ggcacagtcc ctgtacatgg tgggcgtgct gctgggcgcc atggtgtttg gctatctggc
     1141 cgataggctg ggccggagaa aggtgctgat cctgaactac ctgcagacag ccgtgtccgg
     1201 aacctgcgca gccttcgccc ctaattttcc aatctattgt gccttcaggc tgctgtctgg
     1261 aatggcactg gcaggcatca gcctgaactg catgaccctg aatgtggagt ggatgcctat
     1321 ccacacaagg gcatgcgtgg gcaccctgat cggatacgtg tattccctgg gacagtttct
     1381 gctggcagga gtggcatacg cagtgcctca ctggaggcac ctgcagctgc tggtgtctgc
     1441 cccattcttt gccttcttta tctattcttg gttctttatc gagagcgcca gatggcacag
     1501 ctcctctggc cggctggacc tgacactgag agccctgcag cgggtggcca gaatcaacgg
     1561 caagagggag gagggagcaa agctgtccat ggaggtgctg agagcctctc tgcagaagga
     1621 gctgaccatg ggcaagggac aggcaagcgc catggagctg ctgaggtgcc caacactgcg
     1681 ccacctgttc ctgtgcctga gcatgctgtg gttcgccacc tcctttgcct actatggcct
     1741 ggtcatggac ctgcagggct tcggcgtgtc tatctacctg atccaggtca tctttggcgc
     1801 cgtggatctg ccagccaagc tggtgggctt cctggtcatc aacagcctgg gcaggcgccc
     1861 cgcacagatg gccgccctgc tgctggcagg aatctgcatc ctgctgaatg gcgtgatccc
     1921 ccaggatcag tccatcgtga ggacatctct ggccgtgctg ggcaagggat gcctggcagc
     1981 ctccttcaac tgtatctttc tgtacaccgg cgagctgtat cccacaatga tcaggcagac
     2041 cggcatgggc atgggctcta caatggcaag ggtgggaagc atcgtgtccc ctctggtgtc
     2101 catgaccgcc gagctgtacc cctctatgcc tctgttcatc tatggagcag tgccagtggc
     2161 agcatctgcc gtgacagtgc tgctgccaga gaccctggga cagccactgc cagacacagt
     2221 gcaggatctg gagagcagat gggcaccaac ccagaaggag gcaggcatct accctaggaa
     2281 gggcaagcag acccgccagc agcaggagca ccagaagtat atggtgccac tgcaggcatc
     2341 cgcccaggag aagaatggcc tggacccagc tttcttgtac aaagttggca ttataagaaa
     2401 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2461 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2521 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2581 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2641 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2701 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2761 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2821 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2881 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2941 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3001 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3061 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3121 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3181 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3241 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3301 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3361 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3421 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3481 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3541 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3721 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4141 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4201 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
