LOCUS       sequence-333109-        4590 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 4590)
  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 4590)
  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-SLC20A1_STOP
FEATURES             Location/Qualifiers
     source          1..4590
                     /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(2179..2278)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2295..2314)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2296..2314)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2319..2335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2319..2335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2437..2456)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2448..3257
                     /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(2515..2534)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3404..3992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3893..3912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4146..4163
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4322..4349)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4441..4527)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4576..4590,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4590,1..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 gccaccctga tcacatctac cacagcagca accgcagcaa
      181 gcggaccact ggtggactac ctgtggatgc tgatcctggg cttcatcatc gcctttgtgc
      241 tggccttcag cgtgggcgcc aacgatgtgg ccaatagctt cggcaccgca gtgggatccg
      301 gagtggtgac actgaagcag gcctgcatcc tggcctctat ctttgagacc gtgggaagcg
      361 tgctgctggg agcaaaggtg tccgagacaa tccggaaggg cctgatcgac gtggagatgt
      421 ataactctac acagggactg ctgatggcag gaagcgtgtc cgccatgttc ggatccgccg
      481 tgtggcagct ggtggcatcc tttctgaagc tgcctatctc tggcacccac tgtatcgtgg
      541 gcgccacaat cggcttctct ctggtggcca agggccagga gggcgtgaag tggagcgagc
      601 tgatcaagat cgtgatgtct tggttcgtga gccctctgct gtccggcatc atgtctggca
      661 tcctgttctt tctggtgagg gcctttatcc tgcacaaggc agacccagtg cctaacggac
      721 tgcgcgccct gccagtgttc tacgcctgca ccgtgggcat caatctgttt agcatcatgt
      781 atacaggcgc ccctctgctg ggcttcgata agctgccact gtggggcacc atcctgatct
      841 ccgtgggctg cgccgtgttt tgtgccctga tcgtgtggtt cttcgtgtgc ccacggatga
      901 agagaaagat cgagcgggag atcaagtgtt ctcctagcga gtccccactg atggagaaga
      961 agaacagcct gaaggaggac cacgaggaga caaagctgtc cgtgggcgat atcgagaata
     1021 agcaccccgt gtctgaagtg ggaccagcaa ccgtgcctct gcaggccgtg gtggaggaga
     1081 gaacagtgag cttcaagctg ggcgacctgg aggaggcacc agagagggag agactgccat
     1141 ccgtggacct gaaggaggag acctccatcg attctacagt gaacggagca gtgcagctgc
     1201 caaacggaaa tctggtgcag tttagccagg ccgtgtccaa ccagatcaat agctccggcc
     1261 actaccagta tcacaccgtg cacaaggact ctggcctgta caaggagctg ctgcacaagc
     1321 tgcacctggc caaagtgggc gattgcatgg gcgactccgg cgataagcca ctgaggagaa
     1381 acaatagcta cacctcctat acaatggcca tctgtggcat gcctctggat agcttcaggg
     1441 caaaggaggg agagcagaag ggagaggaga tggagaagct gacctggcct aacgccgact
     1501 ccaagaagag gatccgcatg gattcttaca caagctattg caatgccgtg agcgacctgc
     1561 actctgccag cgagatcgat atgtccgtga aggcagagat gggactgggc gacaggaagg
     1621 gatccaatgg ctctctggag gagtggtacg accaggataa gccagaggtg agcctgctgt
     1681 tccagtttct gcagatcctg accgcatgct tcggatcctt tgcccacgga ggcaacgacg
     1741 tgagcaatgc catcggccca ctggtggccc tgtacctggt gtatgacacc ggcgacgtga
     1801 gcagcaaggt ggccacaccc atctggctgc tgctgtatgg aggagtggga atctgcgtgg
     1861 gactgtgggt gtggggcagg cgcgtgatcc agaccatggg caaggacctg accccaatca
     1921 caccctcctc tggcttctcc atcgagctgg cctctgccct gacagtggtc atcgccagca
     1981 acatcggcct gcccatctcc accacacact gtaaagtggg aagcgtggtg tccgtgggat
     2041 ggctgcggag caagaaggca gtggattgga ggctgtttcg caatatcttc atggcctggt
     2101 ttgtgaccgt gcctatctct ggcgtgatca gcgccgccat catggccatc tttcggtacg
     2161 tgatcctgag aatgtaagac ccagctttct tgtacaaagt tggcattata agaaagcatt
     2221 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2281 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2341 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2401 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2461 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2521 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     2581 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2641 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2701 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2761 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2821 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2881 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2941 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3001 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3061 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3121 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3181 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3241 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3301 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3361 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3421 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3481 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3541 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3601 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3661 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3721 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3781 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3841 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3901 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3961 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4021 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4081 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4141 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4201 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4261 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     4321 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     4381 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     4441 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     4501 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
     4561 tagcatggat gttttcccag tcacgacgtt
//
