LOCUS       sequence-365766-        6189 bp    DNA     circular SYN 15-DEC-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 6189)
  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 6189)
  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-SLC12A2_STOP
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3746..3845)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3862..3881)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3863..3881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3886..3902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3886..3902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4004..4023)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4015..4824
                     /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(4082..4101)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4971..5559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5460..5479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5713..5730
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5889..5916)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6008..6094)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6143..6165
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6157..6174
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6158..6174
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgga gccacggccc
      121 acagccccaa gctccggagc accaggactg gcaggagtgg gagagacccc atccgccgcc
      181 gccctggccg ccgccagagt ggagctgccc ggcacagccg tgcccagcgt gcctgaggac
      241 gccgcccctg cctccaggga cggcggcggc gtgcgcgatg agggccctgc cgccgccggc
      301 gacggcctgg gcaggccact gggacctacc ccaagccagt cccgcttcca ggtggatctg
      361 gtgtccgaga acgccggcag agccgccgca gccgccgcag cagcagcagc agccgccgcc
      421 gccgctggag ccggcgccgg cgccaagcag acccccgccg acggcgaggc atctggagag
      481 agcgagcctg caaagggaag cgaggaggca aagggccggt tcagagtgaa ttttgtggat
      541 cctgccgcct ctagctccgc cgaggactct ctgagcgatg cagcaggagt gggagtggac
      601 ggaccaaacg tgtccttcca gaatggcggc gatacagtgc tgtctgaggg ctctagcctg
      661 cactctggcg gaggaggagg aagcggacac caccagcact actattacga cacccacaca
      721 aacacctatt acctgcggac atttggccac aataccatgg acgccgtgcc tcggatcgat
      781 cactacagac acaccgcagc acagctggga gagaagctgc tgagaccaag cctggccgag
      841 ctgcacgacg agctggagaa ggagcccttc gaggatggct ttgccaacgg cgaggagtcc
      901 acacctacca gggatgccgt ggtgacatat accgccgagt ctaagggcgt ggtgaagttc
      961 ggctggatca agggcgtgct ggtgcggtgc atgctgaata tctggggcgt gatgctgttt
     1021 atcagactga gctggatcgt gggacaggca ggaatcggac tgagcgtgct ggtcatcatg
     1081 atggccacag tggtgaccac aatcacaggc ctgtccacct ctgccatcgc aaccaacgga
     1141 ttcgtgcggg gaggaggcgc ctattacctg atcagcagat ccctgggacc agagtttgga
     1201 ggagcaatcg gcctgatctt cgcctttgcc aacgcagtgg cagtggcaat gtacgtggtg
     1261 ggcttcgccg agacagtggt ggagctgctg aaggagcact ccatcctgat gatcgacgag
     1321 atcaatgata tcaggatcat cggcgccatc accgtggtca tcctgctggg catcagcgtg
     1381 gcaggaatgg agtgggaggc aaaggcccag atcgtgctgc tggtcatcct gctgctggcc
     1441 atcggcgact ttgtgatcgg caccttcatc ccactggagt ctaagaagcc caagggcttc
     1501 tttggctaca agagcgagat cttcaacgag aattttggcc ccgatttccg cgaggaggag
     1561 acattcttta gcgtgttcgc catcttcttt cctgcagcaa ccggaatcct ggcaggagca
     1621 aacatctccg gcgacctggc cgatcctcag tctgccatcc caaagggcac actgctggcc
     1681 atcctgatca ccacactggt gtacgtgggc atcgccgtgt ccgtgggatc ttgcgtggtg
     1741 cgggacgcaa ccggaaacgt gaatgataca atcgtgaccg agctgacaaa ctgcaccagc
     1801 gccgcctgta agctgaattt cgacttttcc tcttgcgaga gctccccatg ttcctatggc
     1861 ctgatgaaca attttcaggt catgagcatg gtgtccggct tcacacccct gatcagcgcc
     1921 ggcatctttt ccgccaccct gtctagcgcc ctggcatctc tggtgagcgc cccaaagatc
     1981 ttccaggccc tgtgcaagga taacatctat cccgccttcc agatgtttgc caagggctac
     2041 ggcaagaaca atgagccact gagaggctat atcctgacct ttctgatcgc cctgggcttc
     2101 atcctgatcg ccgagctgaa cgtgatcgcc cccatcatct ccaatttctt tctggcctct
     2161 tacgccctga tcaatttctc cgtgtttcac gcaagcctgg caaagtcccc tggatggagg
     2221 ccagccttca agtattacaa catgtggatc agcctgctgg gcgccatcct gtgctgtatc
     2281 gtgatgtttg tgatcaattg gtgggccgcc ctgctgacat acgtgatcgt gctgggcctg
     2341 tatatctacg tgacctataa gaagcctgac gtgaactggg gctcctctac acaggccctg
     2401 acctatctga atgccctgca gcactctatc aggctgagcg gcgtggagga tcacgtgaag
     2461 aacttccgcc cacagtgcct ggtcatgaca ggagcaccaa attccaggcc tgccctgctg
     2521 cacctggtgc acgactttac caagaacgtg ggcctgatga tctgtggaca cgtgcacatg
     2581 ggaccacgga gacaggccat gaaggagatg agcatcgatc aggccaagta ccagcggtgg
     2641 ctgatcaaga ataagatgaa ggccttctat gcaccagtgc acgcagacga tctgagggag
     2701 ggcgcccagt acctgatgca ggcagcagga ctgggaagga tgaagcctaa caccctggtg
     2761 ctgggcttta agaaggactg gctgcaggcc gatatgaggg acgtggatat gtacatcaat
     2821 ctgtttcacg acgccttcga tatccagtat ggcgtggtgg tcatccgcct gaaggagggc
     2881 ctggacatca gccacctgca gggccaggag gagctgctga gctcccagga gaagtctcca
     2941 ggcaccaagg atgtggtggt gagcgtggag tactccaaga agtctgacct ggatacatcc
     3001 aagcccctgt ctgagaagcc tatcacccac aaggtggagg aggaggacgg caagacagcc
     3061 acccagcccc tgctgaagaa ggagtctaag ggcccaatcg tgcccctgaa cgtggccgac
     3121 cagaagctgc tggaggccag cacacagttc cagaagaagc agggcaagaa taccatcgac
     3181 gtgtggtggc tgtttgacga tggcggcctg accctgctga tcccctacct gctgaccacc
     3241 aagaagaagt ggaaggactg taagatccgg gtgttcatcg gcggcaagat caacagaatc
     3301 gaccacgata ggcgcgccat ggccacactg ctgtctaagt ttagaatcga cttcagcgat
     3361 atcatggtgc tgggcgatat caacaccaag cctaagaagg agaatatcat cgcctttgag
     3421 gagatcatcg agccatatcg gctgcacgag gacgataagg agcaggacat cgccgataag
     3481 atgaaggagg acgagccctg gagaatcaca gataacgagc tggagctgta taagacaaag
     3541 acctacaggc agatccgcct gaacgaactg ctgaaggaac actctagcac cgccaatatc
     3601 atcgtgatgt ccctgcctgt ggcaaggaag ggagccgtgt cctctgccct gtacatggcc
     3661 tggctggagg ccctgtctaa ggacctgcca cctatcctgc tggtgcgcgg caatcaccag
     3721 agcgtgctga ccttctattc ctaagaccca gctttcttgt acaaagttgg cattataaga
     3781 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     3841 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     3901 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     3961 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     4021 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     4081 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     4141 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     4201 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     4261 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     4321 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     4381 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     4441 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     4501 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     4561 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     4621 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     4681 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     4741 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     4801 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     4861 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     4921 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5641 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5701 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5761 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5821 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     5881 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     5941 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     6001 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     6061 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     6121 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     6181 ctcgggccc
//
