LOCUS       sequence-263654-        4323 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 4323)
  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 4323)
  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_SLC19A1
FEATURES             Location/Qualifiers
     source          1..4323
                     /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(2448..2547)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2564..2583)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2565..2583)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2588..2604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2588..2604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2706..2725)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2717..3526
                     /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(2784..2803)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3673..4261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4162..4181
                     /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 accatggtgc ccagctcccc tgcagtggag aagcaggtgc cagtggagcc
      721 aggacctgac ccagagctgc ggtcttggag acacctggtg tgctacctgt gcttctatgg
      781 ctttatggcc cagatcagac ctggcgagag cttcatcacc ccatacctgc tgggccccga
      841 caagaacttt acaagggagc aggtgaccaa tgagatcaca cctgtgctgt cctactctta
      901 tctggccgtg ctggtgccag tgttcctgct gaccgattac ctgcgctata cacccgtgct
      961 gctgctgcag ggcctgagct tcgtgagcgt gtggctgctg ctgctgctgg gacacagcgt
     1021 ggcccacatg cagctgatgg agctgttcta ctccgtgacc atggccgccc ggatcgccta
     1081 ctctagctat atcttttccc tggtgaggcc agcaaggtac cagagggtgg caggatattc
     1141 tagggcagcc gtgctgctgg gcgtgttcac ctcctctgtg ctgggacagc tgctggtgac
     1201 cgtgggacgg gtgagctttt ccacactgaa ctatatcagc ctggccttcc tgacattttc
     1261 cgtggtgctg gccctgttcc tgaagagacc caagaggtcc ctgttcttta acagagacga
     1321 taggggccgc tgcgagacct ctgccagcga gctggagaga atgaatccag gacctggagg
     1381 caagctggga cacgcactga gggtggcatg tggcgactct gtgctggcaa ggatgctgag
     1441 ggagctgggc gatagcctgc ggagaccaca gctgagactg tggtctctgt ggtgggtgtt
     1501 taacagcgcc ggctactatc tggtggtgta ctatgtgcac atcctgtgga atgaggtgga
     1561 ccccaccaca aactccgcca gggtgtacaa tggagcagca gatgcagcaa gcaccctgct
     1621 gggagcaatc acatccttcg ccgcaggctt tgtgaagatc agatgggcca ggtggagcaa
     1681 gctgctgatc gcaggagtga ccgcaacaca ggcaggactg gtgttcctgc tggcacacac
     1741 acgccaccct agctccatct ggctgtgcta cgccgccttc gtgctgtttc ggggcagcta
     1801 tcagttcctg gtgccaatcg ccacctttca gatcgcctct agcctgtcca aggagctgtg
     1861 cgccctggtg ttcggcgtga ataccttctt tgccacaatc gtgaagacca tcatcacctt
     1921 catcgtgagc gacgtgagag gactgggact gcctgtgagg aagcagttcc agctgtactc
     1981 cgtgtatttt ctgatcctgt ctatcatcta cttcctggga gcaatgctgg atggactgag
     2041 gcactgccag cggggccacc acccacgcca gccacctgca cagggactgc ggtccgccgc
     2101 cgaggagaag gccgcccagg ccctgtctgt gcaggataag ggactgggag gactgcagcc
     2161 tgcacagagc ccaccactgt ccccagagga ctctctggga gcagtgggac cagcatctct
     2221 ggagcagagg cagagcgatc cctatctggc acaggcacca gcaccacagg cagcagagtt
     2281 tctgtccccc gtgaccacac cttctccatg caccctgtgc agcgcccagg catccggacc
     2341 agaggcagca gacgagacat gccctcagct ggcagtgcac cctccaggcg tgtctaagct
     2401 gggcctgcag tgtctgccta gcgatggcgt gcagaacgtg aatcaggacc cagctttctt
     2461 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2521 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2581 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2641 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2701 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2761 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2821 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2881 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2941 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3001 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3061 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3121 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3181 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3241 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3301 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3361 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3421 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3481 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3541 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3601 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3661 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3721 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3781 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3841 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3901 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3961 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4021 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4081 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4141 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4201 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4261 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4321 gtt
//
