LOCUS       sequence-259090-        3951 bp    DNA     circular SYN 14-APR-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 3951)
  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 3951)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_SLC17A1
FEATURES             Location/Qualifiers
     source          1..3951
                     /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(2076..2175)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2192..2211)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2193..2211)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2216..2232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2216..2232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2334..2353)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2345..3154
                     /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(2412..2431)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3301..3889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3790..3809
                     /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 accatgcaga tggacaaccg gctgccccct aagaaggtgc ctggcttctg
      721 ctcctttaga tacggcctgt ctttcctggt gcactgctgt aatgtgatca tcacagccca
      781 gcgggcctgt ctgaacctga ccatggtggt catggtgaat tctacagatc cccacggcct
      841 gcctaacacc agcacaaaga agctgctgga caacatcaag aatcctatgt acaattggag
      901 cccagatatc cagggcatca tcctgagctc cacatcctat ggcgtgatca tcatccaggt
      961 gcccgtgggc tacttctctg gcatctatag caccaagaag atgatcggct ttgccctgtg
     1021 cctgtctagc gtgctgagcc tgctgatccc acccgccgcc ggcatcggcg tggcctgggt
     1081 ggtggtgtgc agggcagtgc agggagcagc acagggcatc gtggccacag cccagttcga
     1141 gatctacgtg aagtgggcac ctccactgga gaggggcaga ctgacctcca tgtctacaag
     1201 cggcttcctg ctgggccctt ttatcgtgct gctggtgacc ggcgtgatct gcgagtccct
     1261 gggatggcca atggtgttct atatctttgg agcatgcgga tgtgccgtgt gcctgctgtg
     1321 gttcgtgctg ttttacgacg atccaaagga ccacccctgt atctccatct ctgagaagga
     1381 gtatatcacc tcctctctgg tgcagcaggt gagctcctct agacagagcc tgccaatcaa
     1441 ggccatcctg aagtccctgc ccgtgtgggc aatcagcacc ggctccttca cattcttttg
     1501 gagccacaac atcatgaccc tgtacacacc catgtttatc aattccatgc tgcacgtgaa
     1561 catcaaggag aatggcttcc tgagctccct gccttatctg tttgcatgga tctgcggaaa
     1621 cctggcagga cagctgtccg acttctttct gacaaggaat atcctgtctg tgatcgccgt
     1681 gcgcaagctg ttcaccgcag caggctttct gctgccagca atcttcggcg tgtgcctgcc
     1741 ttacctgtct agcacattct attccatcgt gatctttctg atcctggcag gagcaaccgg
     1801 atctttctgc ctgggcggcg tgtttatcaa cggcctggat atcgccccaa ggtacttcgg
     1861 ctttatcaag gcctgttcta ccctgacagg catgatcgga ggactgatcg caagcaccct
     1921 gacaggcctg atcctgaagc aggaccccga gagcgcctgg ttcaagacct ttatcctgat
     1981 ggccgccatc aatgtgacag gcctgatctt ttatctgatc gtggccaccg ccgagatcca
     2041 ggattgggcc aaggagaagc agcacacccg cctggaccca gctttcttgt acaaagttgg
     2101 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2161 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2221 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2281 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2341 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2401 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2461 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2521 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2581 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2641 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2701 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2761 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     2821 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     2881 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     2941 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3001 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3061 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3121 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3181 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3241 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3301 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3361 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3421 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3481 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3541 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3601 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3661 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3721 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3781 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3841 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3901 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
