LOCUS       sequence-260445-        4704 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 4704)
  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 4704)
  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_SLC5A3
FEATURES             Location/Qualifiers
     source          1..4704
                     /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(2829..2928)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2945..2964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2946..2964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2969..2985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2969..2985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3087..3106)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3098..3907
                     /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(3165..3184)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4054..4642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4543..4562
                     /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 accatgaggg ccgtgctgga cacagccgat atcgccatcg tggccctgta
      721 cttcatcctg gtcatgtgca tcggcttctt tgccatgtgg aagtccaaca ggtctaccgt
      781 gagcggctat tttctggccg gccgcagcat gacatgggtg accatcggcg ccagcctgtt
      841 cgtgtccaac atcggctctg agcactttat cggactggca ggaagcggag cagcatccgg
      901 cttcgccgtg ggcgcctggg agtttaatgc actgctgctg ctgcagctgc tgggatgggt
      961 gttcatcccc atctacatcc gctccggcgt gtacacaatg cctgagtatc tgtctaagcg
     1021 gttcggcggc cacagaatcc aggtgtactt tgccgccctg tccctgatcc tgtatatctt
     1081 caccaagctg agcgtggacc tgtactccgg cgccctgttt atccaggagt ctctgggctg
     1141 gaatctgtat gtgagcgtga tcctgctgat cggcatgaca gccctgctga cagtgaccgg
     1201 aggactggtg gccgtgatct atacagacac cctgcaggcc ctgctgatga tcatcggcgc
     1261 cctgaccctg atgatcatca gcatcatgga gatcggcggc ttcgaggagg tgaagcggag
     1321 atacatgctg gcctctccag atgtgacaag catcctgctg acctataacc tgtctaacac
     1381 caatagctgt aacgtgagcc caaagaagga ggccctgaag atgctgagga accccacaga
     1441 cgaggatgtg ccatggccag gctttatcct gggacagacc ccagcatccg tgtggtactg
     1501 gtgcgcagat caggtcatcg tgcagcgcgt gctggcagca aagaatatcg cccacgccaa
     1561 gggcagcaca ctgatggccg gcttcctgaa gctgctgcca atgtttatca tcgtggtgcc
     1621 cggcatgatc tccaggatcc tgttcaccga cgatatcgcc tgcatcaacc ccgagcactg
     1681 catgctggtg tgcggatcca gggcaggctg ttctaatatc gcctacccta gactggtcat
     1741 gaagctggtg ccagtgggac tgaggggact gatgatggcc gtgatgatcg ccgccctgat
     1801 gtctgacctg gatagcatct tcaactctgc cagcacaatc tttaccctgg acgtgtataa
     1861 gctgatcagg aagtccgcca gctcccgcga gctgatgatc gtgggcagaa tcttcgtggc
     1921 ctttatggtg gtcatctcta tcgcctgggt gcctatcatc gtggagatgc agggcggcca
     1981 gatgtacctg tatatccagg aggtggcaga ttacctgacc ccacctgtgg ccgccctgtt
     2041 cctgctggcc atcttttgga agcggtgtaa cgagcaggga gccttctacg gaggaatggc
     2101 aggatttgtg ctgggagccg tgcggctgat cctggccttc gcctatagag cccccgagtg
     2161 cgaccagcct gataatagac caggcttcat caaggacatc cactacatgt atgtggccac
     2221 aggcctgttt tgggtgacag gcctgatcac cgtgatcgtg agcctgctga caccaccccc
     2281 taccaaggag cagatccgga ccacaacctt ttggtctaag aagaacctgg tggtgaagga
     2341 gaattgcagc cccaaggagg agccttatca gatgcaggag aagagcatcc tgagatgttc
     2401 cgagaacaat gagaccatca accacatcat ccccaatggc aagtccgagg actctatcaa
     2461 gggcctgcag cctgaggatg tgaacctgct ggtgacatgc agggaggagg gaaatcctgt
     2521 ggcatccctg ggacactctg aggcagagac cccagtggat gcctactcta acggacaggc
     2581 cgccctgatg ggagagaagg agaggaagaa ggagacagac gatggcggcc gctattggaa
     2641 gttcatcgac tggttctgcg gctttaagtc caagtctctg agcaagaggt ccctgcgcga
     2701 tctgatggag gaggaggccg tgtgcctgca gatgctggag gagaccagac aggtgaaagt
     2761 gatcctgaac atcggcctgt tcgccgtgtg cagcctgggc atcttcatgt tcgtgtactt
     2821 cagcctggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2881 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2941 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3001 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     3061 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     3121 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3181 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     3241 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3301 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3361 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3421 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3481 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3541 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3601 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3661 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3721 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3781 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3841 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3901 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3961 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     4021 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4081 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4141 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4201 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4261 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4321 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4381 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4441 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4501 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4561 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4621 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4681 ttgctggcct tttgctcaca tgtt
//
