LOCUS       sequence-259080-        4737 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 4737)
  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 4737)
  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_SLC15A2
FEATURES             Location/Qualifiers
     source          1..4737
                     /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(2862..2961)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2978..2997)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2979..2997)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3002..3018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3002..3018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3120..3139)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3131..3940
                     /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(3198..3217)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4087..4675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4576..4595
                     /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 accatgaacc ctttccagaa gaatgagagc aaggagaccc tgttttctcc
      721 agtgagcatc gaggaggtgc cacctaggcc accatctcct ccaaagaagc ccagccctac
      781 aatctgcggc tctaactacc cactgagcat cgccttcatc gtggtgaatg agttctgtga
      841 gcggttcagc tactatggca tgaaggccgt gctgatcctg tatttcctgt actttctgca
      901 ctggaacgag gatacctcta caagcatcta tcacgccttc agctccctgt gctactttac
      961 cccaatcctg ggagcagcaa tcgcagactc ttggctgggc aagttcaaga caatcatcta
     1021 tctgtctctg gtgtacgtgc tgggccacgt gatcaagagc ctgggcgccc tgcctatcct
     1081 gggaggacag gtggtgcaca ccgtgctgtc cctgatcggc ctgtctctga tcgccctggg
     1141 cacaggcggc atcaagccat gcgtggcagc cttcggcggc gatcagtttg aggagaagca
     1201 cgccgaggag cggaccagat atttcagcgt gttttacctg tccatcaacg ccggctccct
     1261 gatctctacc ttcatcacac caatgctgag gggcgacgtg cagtgcttcg gcgaggattg
     1321 ttatgcactg gcctttggcg tgccaggact gctgatggtc atcgccctgg tggtgttcgc
     1381 catgggctcc aagatctaca acaagccccc tccagagggc aatatcgtgg cccaggtgtt
     1441 caagtgcatc tggtttgcca tcagcaaccg gtttaagaat agatccggcg atatccccaa
     1501 gaggcagcac tggctggact gggcagcaga gaagtatcct aagcagctga tcatggatgt
     1561 gaaggccctg acccgggtgc tgttcctgta catcccactg cccatgtttt gggccctgct
     1621 ggaccagcag ggcagcagat ggacactgca ggccatcagg atgaaccgca atctgggctt
     1681 ctttgtgctg cagccagatc agatgcaggt gctgaacccc ctgctggtgc tgatcttcat
     1741 ccctctgttc gactttgtga tctacaggct ggtgtccaag tgcggcatca atttctctag
     1801 cctgcgcaag atggccgtgg gcatgatcct ggcatgtctg gcctttgccg tggcagcagc
     1861 cgtggagatc aagatcaacg agatggcacc agcacagcct ggaccacagg aggtgttcct
     1921 gcaggtgctg aacctggccg acgatgaggt gaaggtgacc gtggtgggca atgagaacaa
     1981 tagcctgctg atcgagagca tcaagtcctt tcagaagacc cctcactatt ctaagctgca
     2041 cctgaagaca aagagccagg atttccactt tcacctgaag tatcacaatc tgtccctgta
     2101 cacagagcac tctgtgcagg agaagaactg gtactccctg gtcatccggg aggatggcaa
     2161 ttctatctcc tctatgatgg tgaaggacac cgagtcccgg accacaaacg gcatgaccac
     2221 agtgagattc gtgaacacac tgcacaagga cgtgaatatc tccctgtcta ccgatacaag
     2281 cctgaatgtg ggcgaggact atggcgtgtc cgcctacagg accgtgcaga ggggagagta
     2341 tccagcagtg cactgtagaa cagaggataa gaacttttct ctgaatctgg gactgctgga
     2401 cttcggagca gcatacctgt ttgtgatcac caacaataca aaccagggcc tgcaggcctg
     2461 gaagatcgag gacatcccag ccaataagat gagcatcgcc tggcagctgc cacagtatgc
     2521 actggtgacc gcaggcgaag tgatgttcag cgtgacaggc ctggagttta gctactccca
     2581 ggcccctagc tccatgaagt ccgtgctgca ggcagcatgg ctgctgacca tcgcagtggg
     2641 caacatcatc gtgctggtgg tggcacagtt cagcggactg gtgcagtggg ccgagttcat
     2701 cctgttttcc tgcctgctgc tggtcatctg tctgatcttt tctatcatgg gctactatta
     2761 cgtgccagtg aagaccgagg acatgagggg accagcagat aagcacatcc ctcacatcca
     2821 gggcaatatg atcaagctgg agaccaagaa gacaaagctg gacccagctt tcttgtacaa
     2881 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2941 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     3001 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     3061 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     3121 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     3181 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     3241 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3301 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3361 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3421 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3481 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3541 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3601 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3661 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3721 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3781 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3841 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3901 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3961 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     4021 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4201 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4381 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4441 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4501 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4561 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4621 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4681 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
