LOCUS       sequence-479328-        3216 bp    DNA     circular SYN 30-JUL-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 3216)
  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 3216)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 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-SLC50A1_STOP
FEATURES             Location/Qualifiers
     source          1..3216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(48..636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(128..147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(783..1592)
                     /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     1506..1525
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1584..1603
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1705..1721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1705..1721
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1726..1745
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1726..1744
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1762..1861
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2534..2633)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2649..2666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2649..2665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2658..2680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2729..2815
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2907..2934
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3093..3110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
       61 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      661 gggtctgacg ctcagtggaa cgacgcgtaa ctcacgttaa gggattttgg tcatgagctt
      721 gcgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct
      781 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
      841 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
      901 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
      961 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1021 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1081 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1141 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1201 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1261 tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca
     1321 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1381 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1441 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
     1501 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1561 ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     1621 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     1681 ttttgagaca cgggccagag ctgccaggaa acagctatga ccatgtaata cgactcacta
     1741 taggggatat cagctggatg gcaaataatg attttatttt gactgatagt gacctgttcg
     1801 ttgcaacaaa ttgataagca atgctttctt ataatgccaa ctttgtacaa gaaagctggg
     1861 tcttaggtct gcagcagcca ataattccga tcctgctcct gggggtactt ccagaacagc
     1921 caaaagcgga tgaagcttgt cacgatgccg gggaagttag acaccatgat atatgggtcg
     1981 cgcagcctaa agccgtacag acaccaggag gcgctggtca gcagtgtggc gatggtcagt
     2041 ggataggaca ggcactgtgt gctcttggtc tggatcactt tggccaggtc ggccagagga
     2101 gacaggtaca tggagatggt gaacacggaa caaaacaggc ccagctgctg cagccttgcc
     2161 tcagggtttg gcaccagcag ccagaaatat ccgtatccca gcagcagcac gcccagcagt
     2221 gtggcggtct gcagcagcac cactctcttc cgagggcaat agtgcaggta ggccaggata
     2281 tacagggtct gcagggcggc gcccactgta ttcaccacga tcaggatgcc atcgcccttc
     2341 agggcgccgt aagacagcca gcccagattg ttcacctctg tggtcagaaa gggcaggaac
     2401 tgcacgttat ccacggacct ggtcattctc atgtgccgca ggtcgctcag gccggcagag
     2461 aacatgccca gtgtaaacac cacgcaggcg ccgtagatca ggctgtccag aaagccgccg
     2521 gcctccatgg tggagcctgc ttttttgtac aaagttggca ttataaaaaa gcattgctca
     2581 tcaatgtgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttggggcccg
     2641 agcttaagac tggccgtcgt tttacaacgt cgtgactggg aaaacatcca tgctagcgtt
     2701 aacgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cggaagactg
     2761 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     2821 gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     2881 ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc tttttgcgtt
     2941 tctacaaact cttcctggct agcggtacgc gtattaattg cgttgcgctc actgcccgct
     3001 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3061 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3121 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3181 tcaggggata acgcaggaaa gaacatgtga gcaaaa
//
