LOCUS       sequence-322152-        6384 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 6384)
  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 6384)
  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-NPC1
FEATURES             Location/Qualifiers
     source          1..6384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3973..4072)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4089..4108)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4090..4108)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4113..4129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4113..4129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4231..4250)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4242..5051
                     /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(4309..4328)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5198..5786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5687..5706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5940..5957
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6116..6143)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6235..6321)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6370..6384,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6384,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacac attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccaccatg accgcaaggg gactggccct gggactgctg ctgctgctgc
      181 tgtgcccagc ccaggtgttc tctcagagct gcgtgtggta tggcgagtgt ggcatcgcct
      241 acggcgacaa gcggtacaac tgcgagtaca gcggaccacc taagccactg ccaaaggacg
      301 gctacgatct ggtgcaggag ctgtgccctg gcttcttttt cggcaacgtg tccctgtgct
      361 gtgacgtgcg gcagctgcag accctgaagg ataatctgca gctgcctctg cagtttctgt
      421 ctagatgccc aagctgtttt tataacctgc tgaatctgtt ctgcgagctg acctgttccc
      481 caaggcagtc tcagttcctg aacgtgaccg ccacagagga ctacgtggat cccgtgacaa
      541 accagaccaa gacaaatgtg aaggagctgc agtactatgt gggccagagc ttcgccaacg
      601 ccatgtataa tgcatgcagg gacgtggagg caccaagctc caatgataag gccctgggac
      661 tgctgtgcgg caaggacgca gatgcctgta acgccaccaa ttggatcgag tacatgttta
      721 acaaggataa tggccaggcc cctttcacca tcacaccagt gttttccgac ttccccgtgc
      781 acggcatgga gcctatgaac aatgccacca agggctgcga cgagtctgtg gatgaggtga
      841 cagccccttg cagctgtcag gattgctcca tcgtgtgcgg acctaagcca cagccaccac
      901 ctccaccagc accatggacc atcctgggac tggacgccat gtatgtgatc atgtggatca
      961 catacatggc ctttctgctg gtgtttttcg gcgccttttt cgccgtgtgg tgctatcgga
     1021 agagatactt cgtgagcgag tacaccccca tcgacagcaa catcgccttc agcgtgaatg
     1081 cctccgacaa gggcgaggca tcctgctgtg atcccgtgag cgccgccttc gagggatgcc
     1141 tgcggagact gtttacccgg tggggctctt tctgcgtgag aaacccaggc tgcgtgatct
     1201 ttttcagcct ggtgtttatc acagcctgtt ctagcggcct ggtgttcgtg agggtgacca
     1261 caaatccagt ggatctgtgg agcgccccat cctctcaggc aaggctggag aaggagtact
     1321 ttgaccagca cttcggccca tttttcagga ccgagcagct gatcatcagg gcaccactga
     1381 cagataagca catctaccag ccatatccta gcggagcaga cgtgcccttc ggccctccac
     1441 tggatatcca gatcctgcac caggtgctgg acctgcagat cgccatcgag aacatcaccg
     1501 cctcctatga caatgagacc gtgacactgc aggatatctg cctggcccct ctgagcccat
     1561 ataacaccaa ttgtacaatc ctgtccgtgc tgaactactt tcagaattcc cactctgtgc
     1621 tggatcacaa gaagggcgac gacttcttcg tgtacgccga ctatcacacc cacttcctgt
     1681 actgcgtgcg ggcccctgcc tctctgaacg acaccagcct gctgcacgat ccatgtctgg
     1741 gcacatttgg cggacccgtg ttcccttggc tggtgctggg cggctatgac gatcagaact
     1801 acaacaatgc caccgccctg gtcatcacct tccctgtgaa caattactat aatgataccg
     1861 agaagctgca gagagcccag gcctgggaga aggagtttat caacttcgtg aagaattaca
     1921 agaacccaaa tctgaccatc agcttcacag ccgagcggtc catcgaggac gagctgaaca
     1981 gagagagcga ctccgacgtg ttcaccgtgg tcatcagcta tgccatcatg ttcctgtaca
     2041 tcagcctggc cctgggccac atgaagtcct gcaggcgcct gctggtggat agcaaggtgt
     2101 ccctgggcat cgccggcatc ctgatcgtgc tgagctccgt ggcctgttcc ctgggcgtgt
     2161 tctcttatat cggcctgcct ctgaccctga tcgtgatcga agtgatccca tttctggtgc
     2221 tggccgtggg cgtggacaat atcttcatcc tggtgcaggc ataccagagg gacgagcgcc
     2281 tgcagggaga gacactggat cagcagctgg gaagggtgct gggagaggtg gcaccatcca
     2341 tgtttctgtc tagcttctct gagaccgtgg cctttttcct gggcgccctg tccgtgatgc
     2401 cagcagtgca cacattttct ctgttcgccg gcctggccgt gtttatcgac ttcctgctgc
     2461 agatcacctg cttcgtgagc ctgctgggcc tggacatcaa gaggcaggag aagaaccgcc
     2521 tggatatctt ctgctgcgtg cggggagcag aggacggcac atccgtgcag gcctctgaga
     2581 gctgtctgtt ccggtttttc aagaattcct attctccact gctgctgaag gactggatga
     2641 gacccatcgt gatcgccatc tttgtgggcg tgctgagctt ctccatcgcc gtgctgaaca
     2701 aggtggacat cggcctggat cagtctctga gcatgcccga cgatagctat atggtggatt
     2761 actttaagtc catctctcag tatctgcacg caggaccacc cgtgtacttc gtgctggagg
     2821 agggccacga ctacacctcc tctaagggcc agaatatggt gtgcggcggc atgggctgta
     2881 acaatgattc cctggtgcag cagatcttca acgccgccca gctggacaat tatacacgga
     2941 tcggctttgc ccccagctcc tggatcgacg attacttcga ttgggtgaag cctcagtcta
     3001 gctgctgtag agtggacaac atcaccgatc agttttgcaa tgcaagcgtg gtggaccctg
     3061 catgcgtgag gtgtaggcca ctgacaccag agggcaagca gaggccacag ggaggcgact
     3121 tcatgcgctt cctgcctatg ttcctgtccg acaacccaaa tccaaagtgt ggcaagggag
     3181 gacacgcagc atactcctct gccgtgaaca tcctgctggg acacggaacc agggtgggag
     3241 ccacatactt catgacctac cacacagtgc tgcagacctc cgccgacttc atcgatgccc
     3301 tgaagaaggc caggctgatc gcctctaacg tgaccgagac aatgggcatc aatggctctg
     3361 cctatcgcgt gtttccctac agcgtgtttt acgtgttcta tgagcagtac ctgaccatca
     3421 tcgacgatac aatctttaac ctgggcgtga gcctgggcgc catcttcctg gtgaccatgg
     3481 tgctgctggg atgcgagctg tggtccgccg tgatcatgtg cgccacaatc gccatggtgc
     3541 tggtgaatat gttcggcgtg atgtggctgt ggggcatcag cctgaacgcc gtgtccctgg
     3601 tgaacctggt catgtcttgc ggcatcagcg tggagttttg tagccacatc accagagcct
     3661 tcacagtgtc catgaaggga tctagggtgg agagagcaga ggaggccctg gcacacatgg
     3721 gaagctccgt gttttctggc atcaccctga caaagttcgg cggcatcgtg gtgctggcct
     3781 ttgccaagag ccagatcttc cagatctttt atttccggat gtacctggca atggtgctgc
     3841 tgggagcaac ccacggcctg atctttctgc ccgtgctgct gtcttatatc ggccctagcg
     3901 tgaacaaggc caagtcctgc gccaccgagg agagatacaa gggcacagag cgggagagac
     3961 tgctgaattt cgacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
     4021 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     4081 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     4141 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     4201 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     4261 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     4321 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     4381 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     4441 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     4501 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     4561 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     4621 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     4681 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     4741 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     4801 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     4861 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4921 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4981 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     5041 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     5101 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     5161 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5221 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5281 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5341 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5401 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5461 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5521 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5581 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5641 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5701 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5761 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5821 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5881 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5941 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6001 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6061 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
     6121 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
     6181 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
     6241 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
     6301 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
     6361 ggatgttttc ccagtcacga cgtt
//
