LOCUS       sequence-334672-        4611 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 4611)
  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 4611)
  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-SLC22A23_STOP
FEATURES             Location/Qualifiers
     source          1..4611
                     /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"
     regulatory      1058..1067
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2168..2267)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2284..2303)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2285..2303)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2308..2324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2308..2324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2426..2445)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2437..3246
                     /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(2504..2523)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3393..3981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3882..3901
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4135..4152
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4311..4338)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4430..4516)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4565..4587
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4579..4596
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4580..4596
                     /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 ccaccatggc catcgatcgg
      121 agaagggagg cagcaggagg aggaccaggc cgccagcctg caccagcaga ggagaacgga
      181 tctctgccac ctggcgacgc cgccgccagc gccccactgg gaggaagggc aggcccagga
      241 ggaggagccg agatccagcc actgccacca ctgcaccctg gaggaggccc ccacccttct
      301 tgctgtagcg ccgcagcagc accttccctg ctgctgctgg actacgatgg ctctgtgctg
      361 ccattcctgg gaggactggg aggaggatat cagaagacac tggtgctgct gacctggatc
      421 cccgccctgt tcatcggctt tagccagttc tctgacagct tcctgctgga tcagcctaac
      481 ttttggtgca ggggagcagg caagggaacc gagctggcag gagtgaccac aaccggaagg
      541 ggaggcgata tgggcaattg gacaagcctg ccaacaaccc ccttcgccac cgcaccatgg
      601 gaggcagcag gaaaccgctc caatagctcc ggagcagacg gaggcgatac ccctccactg
      661 ccaagcccac ctgacaaggg cgataacgcc tccaattgcg actgtagagc ctgggattac
      721 ggcatcaggg ccggcctggt gcagaacgtg gtgtctaagt gggacctggt gtgcgataat
      781 gcctggaagg tgcacatcgc caagttcagc ctgctggtgg gcctgatctt tggctatctg
      841 atcacaggct gtatcgcaga ctgggtgggc cgccggcctg tgctgctgtt ttctatcatc
      901 ttcatcctga tctttggcct gaccgtggcc ctgagcgtga acgtgacaat gttctccacc
      961 ctgagattct ttgagggctt ttgcctggcc ggcatcatcc tgacactgta cgcactgagg
     1021 atcgagctgt gcccaccagg caagcggttc atgatcacca tggtggcatc cttcgtggca
     1081 atggcaggac agtttctgat gccaggactg gccgccctgt gcagggattg gcaggtgctg
     1141 caggccctga tcatctgtcc attcctgctg atgctgctgt attggagcat ctttcccgag
     1201 tccctgcggt ggctgatggc cacacagcag ttcgagagcg ccaagagact gatcctgcac
     1261 tttacccaga agaacaggat gaatcccgag ggcgacatca agggcgtgat ccctgagctg
     1321 gagaaggagc tgtccagaag gccaaagaag gtgtgcatcg tgaaggtggt gggcacaaga
     1381 aacctgtgga agaatatcgt ggtgctgtgc gtgaactccc tgaccggcta cggcatccac
     1441 cactgtttcg ccaggtctat gatgggccac gaggtgaagg tgcccctgct ggagaacttc
     1501 tacgccgatt actatacaac cgcctctatc gccctggtga gctgcctggc catgtgcgtg
     1561 gtggtgagat tcctgggccg gagaggagga ctgctgctgt ttatgatcct gaccgccctg
     1621 gcaagcctgc tgcagctggg actgctgaat ctgatcggca agtactccca gcaccctgac
     1681 tctggcatgt ccgactctgt gaaggataag ttctccatcg ccttttctat cgtgggaatg
     1741 ttcgcatccc acgcagtggg aagcctgtcc gtgttctttt gcgccgagat cacacctacc
     1801 gtgatcagat gtggaggact gggactggtg ctggcatctg ccggatttgg catgctgaca
     1861 gccccaatca tcgagctgca caaccagaag ggctatttcc tgcaccacat catctttgcc
     1921 tgctgtaccc tgatctgcat catctgtatc ctgctgctgc ctgagtctag agatcagaat
     1981 ctgccagaga acatcagcaa tggcgagcac tacacaaggc agcctctgct gccacacaag
     2041 aagggagagc agccactgct gctgaccaac gccgagctga aggactattc cggactgcac
     2101 gatgcagcag cagcaggcga cacactgcct gagggcgcca ccgccaatgg catgaaggcc
     2161 atgtaagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     2221 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     2281 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     2341 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     2401 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     2461 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2521 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     2581 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2641 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2701 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     2761 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     2821 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2881 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2941 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     3001 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     3061 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     3121 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3181 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3241 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     3301 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     3361 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3421 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3481 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3541 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3601 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3661 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3721 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3781 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3841 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3901 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3961 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4021 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4081 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4141 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4201 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4261 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     4321 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     4381 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     4441 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     4501 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     4561 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc c
//
