LOCUS       sequence-345626-        6630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6630)
  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 6630)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2149..2158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4187..4286)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4303..4322)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4304..4322)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4327..4343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4327..4343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4445..4464)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4456..5265
                     /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(4523..4542)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5412..6000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5901..5920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6154..6171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6330..6357)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6449..6535)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6584..6606
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6598..6615
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6599..6615
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc agaggcagga
      121 ctgaggggat ggctgctgtg ggccctgctg ctgcgcctgg cccagagcga gccatacacc
      181 acaatccacc agcccggcta ctgtgccttc tatgacgagt gcggcaagaa ccctgagctg
      241 tctggcagcc tgatgaccct gagcaacgtg tcctgtctgt ctaatacccc agcccggaag
      301 atcacaggcg atcacctgat cctgctgcag aagatctgcc ccagactgta taccggccct
      361 aatacacagg cctgctgtag cgccaagcag ctggtgtccc tggaggcctc cctgtctatc
      421 accaaggccc tgctgacacg gtgtccagcc tgcagcgaca acttcgtgaa tctgcactgt
      481 cacaacacct gctctcccaa tcagagcctg tttatcaacg tgacaagagt ggcacagctg
      541 ggagcaggac agctgcctgc cgtggtggcc tacgaggcct tttatcagca ctctttcgcc
      601 gagcagagct acgatagctg ctcccgggtg agagtgccag cagcagccac cctggcagtg
      661 ggaacaatgt gcggcgtgta tggatccgcc ctgtgcaacg cacagcggtg gctgaatttc
      721 cagggcgaca ccggaaatgg actggcacct ctggatatca catttcacct gctggagcca
      781 ggacaggcag tgggatctgg aatccagccc ctgaacgagg gagtggcaag gtgtaatgag
      841 agccagggcg acgatgtggc cacatgttct tgccaggact gtgcagcaag ctgcccagca
      901 atcgcaaggc cacaggccct ggatagcacc ttctacctgg gacagatgcc aggatccctg
      961 gtgctgatca tcatcctgtg cagcgtgttc gccgtggtga caatcctgct ggtgggcttt
     1021 agggtggcac ctgcacgcga caagtccaag atggtggatc caaagaaggg cacctctctg
     1081 agcgacaagc tgtccttctc tacccacaca ctgctgggcc agttctttca gggatgggga
     1141 acctgggtgg caagctggcc cctgacaatc ctggtgctgt ccgtgatccc tgtggtggcc
     1201 ctggcagcag gactggtgtt taccgagctg accacagacc ctgtggagct gtggtccgcc
     1261 ccaaactctc aggccaggag cgagaaggcc tttcacgatc agcacttcgg ccctttcttt
     1321 cgcaccaacc aggtcatcct gacagcccca aataggagct cctaccgcta tgactctctg
     1381 ctgctgggcc ctaagaactt cagcggcatc ctggacctgg atctgctgct ggagctgctg
     1441 gagctgcagg agaggctgag gcacctgcaa gtgtggagcc cagaggcaca gcggaatatc
     1501 tccctgcagg atatctgcta cgccccactg aaccccgaca atacctctct gtacgattgc
     1561 tgtatcaaca gcctgctgca gtattttcag aacaatagaa cactgctgct gctgaccgcc
     1621 aatcagacac tgatgggcca gaccagccag gtggactgga aggatcactt cctgtactgt
     1681 gccaacgccc ctctgacctt taaggacgga acagccctgg ccctgtcctg catggccgat
     1741 tatggcgccc ccgtgttccc attcctggcc atcggcggct acaagggcaa ggactattcc
     1801 gaggccgagg ccctgatcat gaccttctct ctgaacaatt acccagcagg cgatccaagg
     1861 ctggcacagg caaagctgtg ggaggaggcc tttctggagg agatgcgcgc cttccagcgg
     1921 agaatggccg gcatgtttca ggtgacattc atggccgaga ggagcctgga ggacgagatc
     1981 aaccgcacca cagccgagga tctgccaatc tttgccacct cctacatcgt gatcttcctg
     2041 tatatctccc tggccctggg ctcttactct agctggagca gagtgatggt ggacagcaag
     2101 gccaccctgg gactgggagg agtggcagtg gtgctgggcg ccgtgatggc cgccatgggc
     2161 ttcttttctt acctgggcat ccgctcctct ctggtcatcc tgcaggtggt gcccttcctg
     2221 gtgctgtccg tgggagccga taatatcttt atcttcgtgc tggagtatca gcggctgcca
     2281 aggcgccctg gagagccaag agaggtgcac atcggacggg ccctgggaag agtggcacct
     2341 agcatgctgc tgtgcagcct gtccgaggcc atctgcttct ttctgggcgc cctgacacca
     2401 atgcctgccg tgcggacctt cgccctgaca tccggactgg ccgtgatcct ggactttctg
     2461 ctgcagatgt ctgccttcgt ggccctgctg agcctggact ccaagcggca ggaggcctcc
     2521 agactggacg tgtgctgttg cgtgaagcca caggagctgc cacctccagg acagggagag
     2581 ggcctgctgc tgggcttctt tcagaaggcc tacgccccct ttctgctgca ctggatcacc
     2641 aggggcgtgg tgctgctgct gtttctggcc ctgttcggcg tgtctctgta tagcatgtgc
     2701 cacatcagcg tgggactgga tcaggagctg gccctgccca aggactccta cctgctggat
     2761 tattttctgt tcctgaaccg ctactttgaa gtgggcgccc ccgtgtactt cgtgaccaca
     2821 ctgggctata actttagctc cgaggccggc atgaatgcca tctgttctag cgccggctgc
     2881 aacaattttt ccttcaccca gaagatccag tacgccacag agttcccaga gcagtcttat
     2941 ctggccatcc ccgcctcctc ttgggtggac gacttcatcg actggctgac ccctagctcc
     3001 tgttgcagac tgtacatcag cggccccaac aaggataagt tctgcccttc cacagtgaac
     3061 tctctgaatt gtctgaagaa ttgcatgtcc atcaccatgg gaagcgtgcg gccatccgtg
     3121 gagcagtttc acaagtacct gccctggttc ctgaacgaca gacctaatat caagtgtcca
     3181 aagggcggcc tggccgccta ttctaccagc gtgaacctga catctgacgg ccaggtgctg
     3241 gataccgtgg ccatcctgag cccacggctg gagtactctg gcacaatcag cgcccactgc
     3301 aatctgtatc tgctggactc cacctctaga ttcatggcct accacaagcc cctgaagaac
     3361 agccaggatt ataccgaggc cctgagggca gcaagagagc tggcagccaa tatcaccgca
     3421 gacctgagga aggtgcctgg aacagatcca gcctttgagg tgttccccta caccatcaca
     3481 aacgtgtttt acgagcagta tctgacaatc ctgcccgagg gcctgttcat gctgagcctg
     3541 tgcctggtgc ctacctttgc cgtgtcctgc ctgctgctgg gactggacct gagatccggc
     3601 ctgctgaatc tgctgtctat cgtgatgatc ctggtggata cagtgggctt catggccctg
     3661 tggggcatct cctataacgc cgtgtctctg atcaatctgg tgagcgccgt gggcatgtcc
     3721 gtggagttcg tgtctcacat cacccggtcc tttgccatct ctaccaagcc cacatggctg
     3781 gagagagcca aggaggccac aatctccatg ggatctgccg tgttcgcagg agtggcaatg
     3841 accaacctgc ctggaatcct ggtgctggga ctggcaaagg cacagctgat ccagatcttc
     3901 tttttcaggc tgaatctgct gatcacactg ctgggcctgc tgcacggcct ggtgtttctg
     3961 ccagtgatcc tgagctacgt gggaccagac gtgaaccctg ccctggccct ggagcagaag
     4021 agggcagagg aggcagtggc agccgtgatg gtggcctcct gccccaatca ccctagccgc
     4081 gtgtccaccg ccgataacat ctatgtgaat cacagcttcg agggatccat caagggagca
     4141 ggagcaatca gcaacttcct gcccaacaat ggccgccagt tttgagaccc agctttcttg
     4201 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     4261 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     4321 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     4381 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     4441 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     4501 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     4561 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     4621 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     4681 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     4741 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     4801 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     4861 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     4921 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4981 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     5041 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     5101 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     5161 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     5221 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     5281 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     5341 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     5401 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5461 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5521 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5581 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5641 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5701 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5761 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5821 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5881 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5941 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6001 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6061 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     6301 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     6361 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     6421 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     6481 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     6541 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     6601 aaaacgacgg ccagtcttaa gctcgggccc
//
