LOCUS       sequence-259220-        5847 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 5847)
  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 5847)
  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_SLC12A1
FEATURES             Location/Qualifiers
     source          1..5847
                     /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(3972..4071)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4088..4107)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4089..4107)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4112..4128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4112..4128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4230..4249)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4241..5050
                     /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(4308..4327)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5197..5785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5686..5705
                     /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 accatgagcc tgaacaatag ctccaacgtg ttcctggatt ccgtgccctc
      721 taacaccaat agatttcagg tgtccgtgat caacgagaat cacgagtcta gcgccgccgc
      781 cgacgataat acagatcccc ctcactacga ggagacctcc ttcggcgacg aggcacagaa
      841 gcggctgaga atctctttca ggcccggcaa ccaggagtgc tatgataatt ttctgcagag
      901 cggcgagacc gccaagacag atgcctcttt ccacgcctac gacagccaca ccaacacata
      961 ctatctgcag acatttggcc acaataccat ggacgccgtg cctaagatcg agtactatcg
     1021 caacaccggc tccatctctg gcccaaaagt gaatcggccc tccctgctgg agatccacga
     1081 gcagctggcc aagaacgtgg cagtgacccc atcctctgcc gacagggtgg caaacggcga
     1141 tggaatccct ggcgacgagc aggccgagaa taaggaggac gatcaggccg gcgtggtgaa
     1201 gttcggctgg gtgaagggcg tgctggtgag atgcatgctg aacatctggg gcgtgatgct
     1261 gtttatcagg ctgagctgga tcgtgggaga ggcaggaatc ggactgggcg tgctgatcat
     1321 cctgctgtcc accatggtga catctatcac cggcctgagc acatccgcca tcgccaccaa
     1381 tggcttcgtg agaggcggcg gcgcctacta tctgatctct aggagcctgg gccctgagtt
     1441 tggcggctcc atcggcctga tcttcgcctt tgccaacgca gtggcagtgg caatgtacgt
     1501 ggtgggcttc gccgagaccg tggtggatct gctgaaggag tccgactcta tgatggtgga
     1561 tccaacaaat gacatccgga tcatcggctc tatcaccgtg gtcatcctgc tgggcatcag
     1621 cgtggcagga atggagtggg aggcaaaggc ccaggtcatc ctgctggtca tcctgctgat
     1681 cgccatcgcc aacttcttta tcggcaccgt gatcccttcc aacaatgaga agaagtctag
     1741 aggcttcttt aactatcagg cctctatctt cgccgagaat tttggcccca ggttcaccaa
     1801 gggcgagggc ttctttagcg tgttcgccat cttctttcct gcagcaacag gaatcctggc
     1861 aggagcaaat atcagcggcg acctggagga tcctcaggac gcaatcccaa ggggaaccat
     1921 gctggccatc tttatcacca cagtggccta cctgggagtg gcaatctgcg tgggagcatg
     1981 cgtggtgcgg gatgccacag gcaacatgaa tgacaccatc atctccggca tgaactgcaa
     2041 tggctctgcc gcctgtggcc tgggctatga tttcagcagg tgccgccacg agccatgtca
     2101 gtacggcctg atgaacaatt ttcaggtcat gagcatggtg tccggattcg gaccactgat
     2161 cacagcaggc atcttttccg ccaccctgag ctccgccctg gcatctctgg tgagcgcccc
     2221 aaaggtgttt caggccctgt gcaaggacaa catctacaag gccctgcagt tctttgccaa
     2281 gggctatggc aagaacaatg agcctctgag aggctacatc ctgacctttc tgatcgccat
     2341 ggccttcatc ctgatcgccg agctgaacac aatcgcccca atcatcagca atttctttct
     2401 ggcctcctat gccctgatca atttctcctg ttttcacgcc tcttacgcca agagccctgg
     2461 ctggaggcca gcctacggca tctataacat gtgggtgtct ctgttcggcg ccgtgctgtg
     2521 ctgtgccgtg atgtttgtga tcaattggtg ggccgccgtg atcacatatg tgatcgagtt
     2581 ctttctgtac gtgtatgtga cctgcaagaa gcccgatgtg aactggggct ctagcacaca
     2641 ggccctgtcc tacgtgtctg ccctggataa cgccctggag ctgaccacag tggaggacca
     2701 cgtgaagaat ttccgccctc agtgtatcgt gctgacagga ggaccaatga ccaggcctgc
     2761 cctgctggac atcacacacg ccttcaccaa gaattctggc ctgtgcatct gctgtgaggt
     2821 gtttgtgggc cctagaaagc tgtgcgtgaa ggagatgaac agcggcatgg ccaagaagca
     2881 ggcctggctg atcaagaata agatcaaggc cttctacgca gcagtggcag cagattgctt
     2941 tagggacggc gtgcgcagcc tgctgcaggc atccggcctg ggccgcatga agcctaacac
     3001 cctggtcatc ggctataaga agaattggcg gaaggcccca ctgacagaga tcgagaacta
     3061 cgtgggcatc atccacgatg ccttcgactt tgagatcggc gtggtcatcg tgcggatctc
     3121 tcagggcttc gatatcagcc aggtgctgca ggtgcaggag gagctggagc ggctggagca
     3181 ggagagactg gccctggagg ccaccatcaa ggacaacgag tgtgaggagg agagcggagg
     3241 aatcagggga ctgtttaaga aggccggcaa gctgaatatc accaagacca caccaaagaa
     3301 ggacggctcc atcaacacaa gccagtccat gcacgtgggc gagttcaatc agaagctggt
     3361 ggaggcctct acacagttca agaagaagca ggagaagggc accatcgacg tgtggtggct
     3421 gttcgacgat ggcggcctga cactgctgat cccctatatc ctgaccctga ggaagaagtg
     3481 gaaggactgc aagctgcgca tctacgtggg cggcaagatc aaccggatcg aggaggagaa
     3541 gatcgtgatg gccagcctgc tgtccaagtt tagaatcaag ttcgccgata tccacatcat
     3601 cggcgacatc aacatcaggc caaataagga gagctggaag gtgtttgagg agatgatcga
     3661 gccctatcgc ctgcacgagt cctgtaagga tctgaccaca gccgagaagc tgaagcggga
     3721 gacaccatgg aagatcaccg acgccgagct ggaggccgtg aaggagaaga gctaccggca
     3781 ggtgagactg aacgagctgc tgcaggagca cagcagagcc gccaatctga tcgtgctgtc
     3841 cctgccagtg gcaaggaagg gatctatcag cgacctgctg tatatggcct ggctggagat
     3901 cctgaccaag aacctgccac ccgtgctgct ggtgcggggc aaccacaaga atgtgctgac
     3961 attctactcc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     4021 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     4081 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     4141 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     4201 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     4261 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     4321 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     4381 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     4441 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     4501 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     4561 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     4621 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4681 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4741 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4801 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4861 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4921 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4981 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     5041 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     5101 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     5161 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5221 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5281 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5341 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5401 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5461 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5521 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5581 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5641 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5701 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5761 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5821 cttttgctgg ccttttgctc acatgtt
//
