LOCUS       sequence-263895-        4776 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 4776)
  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 4776)
  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_SV2A
FEATURES             Location/Qualifiers
     source          1..4776
                     /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(2901..3000)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3017..3036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3018..3036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3041..3057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3041..3057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3159..3178)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3170..3979
                     /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(3237..3256)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4126..4714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4615..4634
                     /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 accatggagg agggcttccg ggatagagcc gcctttatca ggggcgccaa
      721 ggacatcgcc aaggaggtga agaagcacgc cgccaagaag gtggtgaagg gcctggatcg
      781 ggtgcaggac gagtactctc ggagatccta ttctagattc gaggaggagg acgatgacga
      841 tgactttcca gcccccagcg atggctacta tcgcggcgag ggcacacagg acgaggagga
      901 gggaggagca agctccgatg caaccgaggg acacgacgag gatgacgaga tctacgaggg
      961 agagtatcag ggaatcccaa gggcagagtc cggaggcaag ggcgagagaa tggccgatgg
     1021 agcacctctg gccggcgtga ggggaggact gtctgacggc gagggccccc ctggcggcag
     1081 gggcgaggcc cagaggcgca aggagaggga ggagctggca cagcagtacg aggcaatcct
     1141 gagggagtgc ggacacggcc gcttccagtg gacactgtat tttgtgctgg gactggccct
     1201 gatggcagat ggagtggagg tgtttgtggt gggattcgtg ctgccaagcg ccgagaagga
     1261 tatgtgcctg agcgactcca acaagggcat gctgggactc atcgtctatc ttggtatgat
     1321 ggtgggcgcc ttcctgtggg gaggactggc agacaggctg ggccggagac agtgcctgct
     1381 gatctctctg agcgtgaatt ccgtgttcgc cttcttttct agctttgtgc agggctacgg
     1441 caccttcctg ttttgtaggc tgctgtccgg agtgggaatc ggaggatcta tccctatcgt
     1501 gttctcctat ttttctgagt tcctggccca ggagaagaga ggcgagcacc tgagctggct
     1561 gtgcatgttc tggatgatcg gaggcgtgta tgcagcagca atggcatggg caatcatccc
     1621 acactacggc tggagctttc agatgggctc cgcctatcag ttccactctt ggcgggtgtt
     1681 cgtgctggtg tgcgcctttc caagcgtgtt cgcaatcggc gccctgacca cacagcctga
     1741 gtccccacgc ttctttctgg agaacggcaa gcacgatgag gcctggatgg tgctgaagca
     1801 ggtgcacgac accaatatgc gggccaaggg ccaccctgag agagtgttca gcgtgaccca
     1861 catcaagaca atccaccagg aggatgagct gatcgagatc cagtccgaca ccggcacatg
     1921 gtaccagcgg tggggcgtga gagccctgag cctgggagga caagtgtggg gaaactttct
     1981 gtcctgcttc ggcccagagt ataggcgcat caccctgatg atgatgggcg tgtggtttac
     2041 aatgtccttc tcttactatg gcctgaccgt gtggttccca gatatgatca gacacctgca
     2101 ggccgtggac tacgcctcta ggaccaaggt gtttccagga gagagggtgg agcacgtgac
     2161 cttcaacttt acactggaga atcagatcca caggggcggc cagtacttta atgataagtt
     2221 catcggcctg cgcctgaaga gcgtgtcctt tgaggattcc ctgttcgagg agtgctattt
     2281 tgaggacgtg acctcctcta acacattctt tcggaattgt accttcatca acacagtgtt
     2341 ttacaacaca gatctgttcg agtacaagtt cgtgaactct agactgatca atagcacctt
     2401 cctgcacaac aaggagggat gtccactgga cgtgaccgga acaggagagg gagcctacat
     2461 ggtgtatttc gtgagcttcc tgggcaccct ggccgtgctg cctggcaata tcgtgtctgc
     2521 cctgctgatg gacaagatcg gcaggctgag gatgctggca ggaagctccg tgatgagctg
     2581 cgtgtcctgt ttctttctga gcttcggcaa ctctgagagc gccatgatcg ccctgctgtg
     2641 cctgtttggc ggcgtgtcta tcgccagctg gaatgccctg gatgtgctga cagtggagct
     2701 gtacccctcc gacaagcgga ccacagcctt cggctttctg aacgcactgt gcaagctggc
     2761 agccgtgctg ggcatctcca tctttacctc tttcgtgggc atcacaaagg cagcacctat
     2821 cctgttcgca agcgccgccc tggccctggg ctctagcctg gccctgaagc tgccagagac
     2881 cagaggccag gtgctgcagg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2941 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3001 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     3061 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3121 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3181 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3241 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     3301 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3361 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3421 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3481 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3541 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3601 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3661 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3721 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3781 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3841 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3901 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3961 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     4021 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     4081 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4141 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4201 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4261 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4321 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4381 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4441 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4501 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4561 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4621 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4681 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4741 gttcctggcc ttttgctggc cttttgctca catgtt
//
