LOCUS       sequence-259023-        4674 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 4674)
  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 4674)
  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_SLC15A1
FEATURES             Location/Qualifiers
     source          1..4674
                     /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(2799..2898)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2915..2934)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2916..2934)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2939..2955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2939..2955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3057..3076)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3068..3877
                     /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(3135..3154)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4024..4612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4513..4532
                     /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 accatgggca tgtctaagag ccactccttc tttggctacc ctctgtctat
      721 cttctttatc gtggtgaacg agttctgcga gcggttcagc tactatggca tgagggccat
      781 cctgatcctg tatttcacca actttatcag ctgggacgat aatctgtcca ccgccatcta
      841 ccacaccttc gtggccctgt gctatctgac cccaatcctg ggcgccctga tcgcagacag
      901 ctggctgggc aagtttaaga caatcgtgtc tctgagcatc gtgtacacca tcggccaggc
      961 cgtgacatcc gtgagctcca tcaacgacct gaccgatcac aatcacgacg gcacaccaga
     1021 ttccctgccc gtgcacgtgg tgctgtctct gatcggactg gccctgatcg ccctgggaac
     1081 cggaggcatc aagccatgcg tgtccgcctt cggcggcgat cagtttgagg agggccagga
     1141 gaagcagcgg aacagattct tttctatctt ctatctggcc atcaatgccg gcagcctgct
     1201 gtccaccatc atcacaccca tgctgagggt gcagcagtgc ggcatccact ccaagcaggc
     1261 ctgttaccct ctggccttcg gcgtgccagc cgccctgatg gcagtggccc tgatcgtgtt
     1321 cgtgctgggc tctggcatgt ataagaagtt taagccccag ggcaacatca tgggcaaggt
     1381 ggccaagtgt atcggcttcg ccatcaagaa taggtttcgc caccggtcta aggccttccc
     1441 taagagagag cactggctgg actgggccaa ggagaagtac gatgagcgcc tgatcagcca
     1501 gatcaagatg gtgaccagag tgatgtttct gtatatccca ctgcccatgt tctgggccct
     1561 gtttgaccag cagggaagca ggtggaccct gcaggcaacc acaatgtccg gcaagatcgg
     1621 cgccctggag atccagcccg accagatgca gacagtgaac gccatcctga tcgtgatcat
     1681 ggtgcccatc ttcgatgccg tgctgtaccc tctgatcgcc aagtgcggct tcaacttcac
     1741 ctccctgaag aagatggccg tgggcatggt gctggcctct atggccttcg tggtggccgc
     1801 catcgtgcag gtggagatcg ataagacact gcccgtgttc ccaaagggca acgaggtgca
     1861 gatcaaggtg ctgaacatcg gcaacaatac catgaatatc agcctgccag gcgagatggt
     1921 gacactgggc cccatgtccc agaccaatgc cttcatgaca tttgacgtga acaagctgac
     1981 cagaatcaat atctctagcc caggatcccc tgtgaccgca gtgacagacg atttcaagca
     2041 gggccagagg cacacactgc tggtgtgggc ccctaaccac taccaggtgg tgaaggatgg
     2101 cctgaaccag aagccagaga agggcgagaa tggcatccgc ttcgtgaaca cctttaatga
     2161 gctgatcacc atcacaatgt ccggcaaggt gtatgccaac atctcctctt acaatgcctc
     2221 tacatatcag ttctttccca gcggcatcaa gggcttcacc atcagctcca cagagatccc
     2281 ccctcagtgc cagcctaact tcaatacctt ttacctggag tttggcagcg cctacacata
     2341 tatcgtgcag cggaagaatg actcttgtcc agaggtgaag gtgttcgagg atatcagcgc
     2401 caacaccgtg aatatggccc tgcagatccc ccagtacttt ctgctgacct gcggcgaggt
     2461 ggtgttctcc gtgacaggcc tggagttttc ctattctcag gccccttcta acatgaagag
     2521 cgtgctgcag gcaggatggc tgctgaccgt ggcagtgggc aatatcatcg tgctgatcgt
     2581 ggcaggagca ggacagttca gcaagcagtg ggccgagtac atcctgtttg ccgccctgct
     2641 gctggtggtg tgcgtgatct tcgccatcat ggcccgcttt tacacatata tcaaccctgc
     2701 cgagatcgag gcccagttcg acgaggatga gaagaagaac cggctggaga agagcaatcc
     2761 atacttcatg tccggcgcca attctcagaa gcagatggac ccagctttct tgtacaaagt
     2821 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2881 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2941 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     3001 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     3061 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     3121 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     3181 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     3241 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     3301 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     3361 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     3421 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     3481 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     3541 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3601 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3661 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3721 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3781 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3841 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3901 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3961 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4021 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4081 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4141 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4201 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4261 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4321 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4381 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4441 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4501 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4561 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4621 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
