LOCUS       sequence-334660-        5970 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 5970)
  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 5970)
  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-SLC12A5_STOP
FEATURES             Location/Qualifiers
     source          1..5970
                     /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(3527..3626)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3643..3662)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3644..3662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3667..3683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3667..3683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3785..3804)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3796..4605
                     /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(3863..3882)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4752..5340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5241..5260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5494..5511
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5670..5697)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5789..5875)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5924..5946
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5938..5955
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5939..5955
                     /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 ccaccatgtc tcggagattc
      121 accgtgacaa gcctgccacc tgcaggacca gcacgctctc cagacccaga gtccaggcgc
      181 cactctgtgg cagatccaag gcacctgcca ggagaggacg tgaagggcga tggcaaccca
      241 aaggagagct cccccttcat caatagcacc gacacagaga agggcaagga gtatgatggc
      301 aagaacatgg ccctgtttga ggaggagatg gacacatccc caatggtgtc tagcctgctg
      361 tctggcctgg ccaactacac caatctgcca cagggaagca gagagcacga ggaggccgag
      421 aacaatgagg gcggcaagaa gaagcctgtg caggccccaa ggatgggcac attcatgggc
      481 gtgtacctgc cttgcctgca gaacatcttc ggcgtgatcc tgtttctgag gctgacctgg
      541 gtggtgggaa tcgcaggcat catggagagc ttctgcatgg tgtttatctg ctgttcctgt
      601 accatgctga cagccatctc tatgagcgcc atcgcaacaa atggagtggt gccagcagga
      661 ggatcctact atatgatctc ccggtctctg ggacctgagt tcggaggagc agtgggactg
      721 tgcttctatc tgggcaccac atttgccggc gccatgtaca tcctgggcac catcgagatc
      781 ctgctggcct atctgttccc agccatggcc atctttaagg cagaggacgc aagcggagag
      841 gcagcagcca tgctgaacaa tatgagagtg tatggcacct gcgtgctgac atgtatggcc
      901 accgtggtgt ttgtgggcgt gaagtacgtg aataagttcg ccctggtgtt tctgggctgc
      961 gtgatcctga gcatcctggc catctacgcc ggcgtgatca agtccgcctt cgacccaccc
     1021 aactttccaa tctgcctgct gggcaataga accctgtcca ggcacggctt cgacgtgtgc
     1081 gcaaagctgg catgggaggg caacgagaca gtgaccacaa gactgtgggg cctgttctgc
     1141 tcctctaggt ttctgaatgc cacctgtgat gagtacttca ccagaaacaa tgtgaccgag
     1201 atccagggaa tcccaggagc agcaagcgga ctgatcaagg agaacctgtg gagctcctac
     1261 ctgacaaagg gcgtgatcgt ggagaggagc ggaatgacat ccgtgggact ggcagacgga
     1321 acccctatcg acatggatca cccatacgtg ttctctgata tgacaagcta cttcaccctg
     1381 ctggtgggca tctacttccc atccgtgacc ggaatcatgg caggatctaa tagaagcggc
     1441 gacctgaggg atgcccagaa gagcatccct accggcacaa tcctggccat cgccaccaca
     1501 tccgccgtgt atatctctag cgtggtgctg ttcggagcat gcatcgaggg agtggtgctg
     1561 agagacaagt ttggcgaggc cgtgaacggc aatctggtgg tgggcaccct ggcatggcct
     1621 agcccatggg tcatcgtgat cggctccttc ttttctacat gtggagcagg actgcagtcc
     1681 ctgaccggag cacctaggct gctgcaggca atctctaggg acggaatcgt gccattcctg
     1741 caggtgtttg gccacggcaa ggccaacggc gagccaacat gggccctgct gctgaccgcc
     1801 tgcatctgtg agatcggcat cctgatcgcc tctctggatg aggtggcccc catcctgagc
     1861 atgttctttc tgatgtgcta catgttcgtg aacctggcct gtgccgtgca gacactgctg
     1921 aggaccccca attggaggcc tcgcttccgg tactatcact ggaccctgtc ctttctgggc
     1981 atgtctctgt gcctggccct gatgtttatc tgtagctggt actatgcact ggtggcaatg
     2041 ctgatcgcag gactgatcta caagtatatc gagtatagag gcgccgagaa ggagtggggc
     2101 gatggcatca ggggcctgag cctgtccgcc gcaagatacg cactgctgag gctggaggag
     2161 ggacctccac acacaaagaa ctggaggcct cagctgctgg tgctggtgcg ggtggaccag
     2221 gatcagaatg tggtgcaccc acagctgctg agcctgacct cccagctgaa ggccggcaag
     2281 ggcctgacaa tcgtgggctc tgtgctggag ggcaccttcc tggagaacca cccacaggca
     2341 cagagggcag aggagagcat ccggagactg atggaggccg agaaggtgaa gggcttttgc
     2401 caggtggtca tctcctctaa tctgcgggac ggcgtgagcc acctgatcca gtccggagga
     2461 ctgggaggac tgcagcacaa cacagtgctg gtgggctggc ctcgcaattg gcggcagaag
     2521 gaggatcacc agacctggag aaacttcatc gagctggtga gggagaccac agcaggacac
     2581 ctggccctgc tggtgaccaa gaacgtgtcc atgttccctg gcaatccaga gcggttcagc
     2641 gagggcagca tcgacgtgtg gtggatcgtg cacgatggcg gcatgctgat gctgctgccc
     2701 ttcctgctgc ggcaccacaa agtgtggaga aagtgtaaga tgaggatctt tacagtggcc
     2761 cagatggacg ataatagcat ccagatgaag aaggacctga ccacatttct gtaccacctg
     2821 cgcatcaccg ccgaggtgga ggtggtggag atgcacgagt ccgatatctc tgcctacaca
     2881 tatgagaaga ccctggtcat ggagcagaga tcccagatcc tgaagcagat gcacctgaca
     2941 aagaacgaga gagagaggga gatccagtct atcaccgacg agagcagggg ctccatcagg
     3001 cgcaagaacc cagccaatac acgcctgcgg ctgaatgtgc cagaggagac cgcaggcgac
     3061 agcgaggaga agcctgagga ggaggtgcag ctgatccacg atcagtctgc cccaagctgc
     3121 cctagctcct ctccatcccc tggagaggag ccagagggag agggagagac cgaccctgag
     3181 aaggtgcacc tgacctggac aaaggataag tccgtggccg agaagaacaa gggcccatct
     3241 cccgtgagct ccgagggcat caaggacttc ttttctatga agcctgagtg ggagaacctg
     3301 aatcagagca acgtgcggag aatgcacacc gccgtgcgcc tgaatgaagt gatcgtgaag
     3361 aagtcccggg atgccaagct ggtgctgctg aacatgccag gcccccctag aaacaggaat
     3421 ggcgacgaga attatatgga gttcctggag gtgctgacag agcacctgga tagagtgatg
     3481 ctggtgcgcg gaggaggaag agaagtgatc accatctact cttaagaccc agctttcttg
     3541 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     3601 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     3661 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     3721 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     3781 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     3841 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     3901 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     3961 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     4021 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     4081 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     4141 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     4201 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     4261 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4321 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     4381 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     4441 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     4501 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     4561 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     4621 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     4681 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     4741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4861 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5401 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5461 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5521 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5581 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     5641 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     5701 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     5761 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     5821 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     5881 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     5941 aaaacgacgg ccagtcttaa gctcgggccc
//
