LOCUS       sequence-262975-        6627 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 6627)
  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 6627)
  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-NPC1L1
FEATURES             Location/Qualifiers
     source          1..6627
                     /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"
     regulatory      2717..2726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4752..4851)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4868..4887)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4869..4887)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4892..4908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4892..4908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5010..5029)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5021..5830
                     /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(5088..5107)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5977..6565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6466..6485
                     /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 accatggcag aggcaggact gaggggatgg ctgctgtggg ccctgctgct
      721 gcgcctggcc cagagcgagc catacaccac aatccaccag cccggctact gtgccttcta
      781 tgacgagtgc ggcaagaacc ctgagctgtc tggcagcctg atgaccctga gcaacgtgtc
      841 ctgtctgtct aataccccag cccggaagat cacaggcgat cacctgatcc tgctgcagaa
      901 gatctgcccc agactgtata ccggccctaa tacacaggcc tgctgtagcg ccaagcagct
      961 ggtgtccctg gaggcctccc tgtctatcac caaggccctg ctgacacggt gtccagcctg
     1021 cagcgacaac ttcgtgaatc tgcactgtca caacacctgc tctcccaatc agagcctgtt
     1081 tatcaacgtg acaagagtgg cacagctggg agcaggacag ctgcctgccg tggtggccta
     1141 cgaggccttt tatcagcact ctttcgccga gcagagctac gatagctgct cccgggtgag
     1201 agtgccagca gcagccaccc tggcagtggg aacaatgtgc ggcgtgtatg gatccgccct
     1261 gtgcaacgca cagcggtggc tgaatttcca gggcgacacc ggaaatggac tggcacctct
     1321 ggatatcaca tttcacctgc tggagccagg acaggcagtg ggatctggaa tccagcccct
     1381 gaacgaggga gtggcaaggt gtaatgagag ccagggcgac gatgtggcca catgttcttg
     1441 ccaggactgt gcagcaagct gcccagcaat cgcaaggcca caggccctgg atagcacctt
     1501 ctacctggga cagatgccag gatccctggt gctgatcatc atcctgtgca gcgtgttcgc
     1561 cgtggtgaca atcctgctgg tgggctttag ggtggcacct gcacgcgaca agtccaagat
     1621 ggtggatcca aagaagggca cctctctgag cgacaagctg tccttctcta cccacacact
     1681 gctgggccag ttctttcagg gatggggaac ctgggtggca agctggcccc tgacaatcct
     1741 ggtgctgtcc gtgatccctg tggtggccct ggcagcagga ctggtgttta ccgagctgac
     1801 cacagaccct gtggagctgt ggtccgcccc aaactctcag gccaggagcg agaaggcctt
     1861 tcacgatcag cacttcggcc ctttctttcg caccaaccag gtcatcctga cagccccaaa
     1921 taggagctcc taccgctatg actctctgct gctgggccct aagaacttca gcggcatcct
     1981 ggacctggat ctgctgctgg agctgctgga gctgcaggag aggctgaggc acctgcaagt
     2041 gtggagccca gaggcacagc ggaatatctc cctgcaggat atctgctacg ccccactgaa
     2101 ccccgacaat acctctctgt acgattgctg tatcaacagc ctgctgcagt attttcagaa
     2161 caatagaaca ctgctgctgc tgaccgccaa tcagacactg atgggccaga ccagccaggt
     2221 ggactggaag gatcacttcc tgtactgtgc caacgcccct ctgaccttta aggacggaac
     2281 agccctggcc ctgtcctgca tggccgatta tggcgccccc gtgttcccat tcctggccat
     2341 cggcggctac aagggcaagg actattccga ggccgaggcc ctgatcatga ccttctctct
     2401 gaacaattac ccagcaggcg atccaaggct ggcacaggca aagctgtggg aggaggcctt
     2461 tctggaggag atgcgcgcct tccagcggag aatggccggc atgtttcagg tgacattcat
     2521 ggccgagagg agcctggagg acgagatcaa ccgcaccaca gccgaggatc tgccaatctt
     2581 tgccacctcc tacatcgtga tcttcctgta tatctccctg gccctgggct cttactctag
     2641 ctggagcaga gtgatggtgg acagcaaggc caccctggga ctgggaggag tggcagtggt
     2701 gctgggcgcc gtgatggccg ccatgggctt cttttcttac ctgggcatcc gctcctctct
     2761 ggtcatcctg caggtggtgc ccttcctggt gctgtccgtg ggagccgata atatctttat
     2821 cttcgtgctg gagtatcagc ggctgccaag gcgccctgga gagccaagag aggtgcacat
     2881 cggacgggcc ctgggaagag tggcacctag catgctgctg tgcagcctgt ccgaggccat
     2941 ctgcttcttt ctgggcgccc tgacaccaat gcctgccgtg cggaccttcg ccctgacatc
     3001 cggactggcc gtgatcctgg actttctgct gcagatgtct gccttcgtgg ccctgctgag
     3061 cctggactcc aagcggcagg aggcctccag actggacgtg tgctgttgcg tgaagccaca
     3121 ggagctgcca cctccaggac agggagaggg cctgctgctg ggcttctttc agaaggccta
     3181 cgcccccttt ctgctgcact ggatcaccag gggcgtggtg ctgctgctgt ttctggccct
     3241 gttcggcgtg tctctgtata gcatgtgcca catcagcgtg ggactggatc aggagctggc
     3301 cctgcccaag gactcctacc tgctggatta ttttctgttc ctgaaccgct actttgaagt
     3361 gggcgccccc gtgtacttcg tgaccacact gggctataac tttagctccg aggccggcat
     3421 gaatgccatc tgttctagcg ccggctgcaa caatttttcc ttcacccaga agatccagta
     3481 cgccacagag ttcccagagc agtcttatct ggccatcccc gcctcctctt gggtggacga
     3541 cttcatcgac tggctgaccc ctagctcctg ttgcagactg tacatcagcg gccccaacaa
     3601 ggataagttc tgcccttcca cagtgaactc tctgaattgt ctgaagaatt gcatgtccat
     3661 caccatggga agcgtgcggc catccgtgga gcagtttcac aagtacctgc cctggttcct
     3721 gaacgacaga cctaatatca agtgtccaaa gggcggcctg gccgcctatt ctaccagcgt
     3781 gaacctgaca tctgacggcc aggtgctgga taccgtggcc atcctgagcc cacggctgga
     3841 gtactctggc acaatcagcg cccactgcaa tctgtatctg ctggactcca cctctagatt
     3901 catggcctac cacaagcccc tgaagaacag ccaggattat accgaggccc tgagggcagc
     3961 aagagagctg gcagccaata tcaccgcaga cctgaggaag gtgcctggaa cagatccagc
     4021 ctttgaggtg ttcccctaca ccatcacaaa cgtgttttac gagcagtatc tgacaatcct
     4081 gcccgagggc ctgttcatgc tgagcctgtg cctggtgcct acctttgccg tgtcctgcct
     4141 gctgctggga ctggacctga gatccggcct gctgaatctg ctgtctatcg tgatgatcct
     4201 ggtggataca gtgggcttca tggccctgtg gggcatctcc tataacgccg tgtctctgat
     4261 caatctggtg agcgccgtgg gcatgtccgt ggagttcgtg tctcacatca cccggtcctt
     4321 tgccatctct accaagccca catggctgga gagagccaag gaggccacaa tctccatggg
     4381 atctgccgtg ttcgcaggag tggcaatgac caacctgcct ggaatcctgg tgctgggact
     4441 ggcaaaggca cagctgatcc agatcttctt tttcaggctg aatctgctga tcacactgct
     4501 gggcctgctg cacggcctgg tgtttctgcc agtgatcctg agctacgtgg gaccagacgt
     4561 gaaccctgcc ctggccctgg agcagaagag ggcagaggag gcagtggcag ccgtgatggt
     4621 ggcctcctgc cccaatcacc ctagccgcgt gtccaccgcc gataacatct atgtgaatca
     4681 cagcttcgag ggatccatca agggagcagg agcaatcagc aacttcctgc ccaacaatgg
     4741 ccgccagttt gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     4801 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     4861 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     4921 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     4981 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     5041 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     5101 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     5161 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     5221 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     5281 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     5341 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     5401 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     5461 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     5521 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     5581 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     5641 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     5701 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     5761 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     5821 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     5881 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     5941 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6001 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6061 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6121 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6601 cttttgctgg ccttttgctc acatgtt
//
