LOCUS       sequence-479321-        3978 bp    DNA     circular SYN 25-JUL-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 3978)
  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 3978)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 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-SLC46A2_STOP
FEATURES             Location/Qualifiers
     source          1..3978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(919..1728)
                     /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     1642..1661
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1720..1739
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1841..1857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1841..1857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1862..1881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1862..1880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1898..1997
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3432..3531)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3547..3564)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3547..3563)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3556..3578)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3627..3713
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3805..3832
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga
      841 ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa
      901 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      961 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1021 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1081 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     1141 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     1201 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     1261 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     1321 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1381 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt
     1441 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1501 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1561 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     1621 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1681 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt
     1741 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     1801 aatgtaacat cagagatttt gagacacggg ccagagctgc caggaaacag ctatgaccat
     1861 gtaatacgac tcactatagg ggatatcagc tggatggcaa ataatgattt tattttgact
     1921 gatagtgacc tgttcgttgc aacaaattga taagcaatgc tttcttataa tgccaacttt
     1981 gtacaagaaa gctgggtctt acttctcgat gatatcgcca tatgggctca gaggcacctg
     2041 cttgtaggcc acgatggaga tggggatgat ggccaggaag ctcagaaagc tagacagggc
     2101 gaaacaggag cccacaaaca tgtccatggt cagctgatag atcttattgt acagtgtaga
     2161 ggtcaccacg cctgtcaggg ccagggacag ctgcaggatc acaaacacct tgccgtagga
     2221 gctgcccttg atcagcttag acatggcgga tctgattgtg gtcactggga tcagggcgaa
     2281 cagcatcacg gcccgggcga tatagaacat gtaggtctcc ttcacaaagg ccagcagcag
     2341 ggcgccgctg ccaaaagaca ccatgccgat catgatcatt gtggtatcgc ggaagcacct
     2401 agaaaacacc agcacgccca ggaaggaggt gataaagatt gtatagcctg ctgccattcc
     2461 gtatcccact tgcacctggt tccatcccag tggctccctc agcacgaaca gtgggatcac
     2521 atccactgtg cccaccacgg ccaggtcata gatgatggcg cccacaaaca gcagggcgat
     2581 tgtggtcttg tgaggcttgg ccttgcctgg agaaggtggg tgtcccactg catactgctg
     2641 atccagctga tctgggtcca gggtccggta tgttcccacg gttccggaca cggtgtccac
     2701 tgctggcagc tcctgggaag gcttggccac gctctcaggc accttcagca ccagcaggct
     2761 gtacagcagg gcgaaagagg cacaagacac ggagcaggct gtcaggatca gtccctgtcc
     2821 gctgtgtcct gccatctgct tgaacaggtg gccggaggcc atgctgccac aaaagccggc
     2881 caggcccagc atcaggtcga tcaggatcag gcgcacgctg cgcctgccct cagaggatcc
     2941 cagagatccc agtgccatca cgccagacca gaaggcggaa aagccgccga acagtccatt
     3001 cagggcggct gctccgtaca gcacctccac aggccaatcc agcagcacct tcagcagcag
     3061 gcccagcctg ctcagcagaa agcccagcag gctcatgcag atagagatct ttctgtgata
     3121 ccggtcagac agccatccca gtccgtaggc ggacagcagt gggctcagtc ccaccaccag
     3181 attatagatg atgtagaagt tgctgatggc gcgctgctgc tgatcctcca gggcgcccct
     3241 tgggcttgga gaggcggagt ggttgctaga tcctccggtt ccataggatg ccttcaccac
     3301 cagcagcagt cctgcgtcgt acaggcttgc tgccacctgg gagcttgcca ccacaggctc
     3361 cacccaggtt ctagggtgga accttggcag gtgtcctctc cttgggcatg tcacctctgg
     3421 ggacatggtg gagcctgctt ttttgtacaa agttggcatt ataaaaaagc attgctcatc
     3481 aatgtgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt ggggcccgag
     3541 cttaagactg gccgtcgttt tacaacgtcg tgactgggaa aacatccatg ctagcgttaa
     3601 cgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg
     3661 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     3721 gagcggattt gaacgttgtg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     3781 aaactgccag gcatcaaact aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     3841 tacaaactct tcctggctag cggtacgcgt attaattgcg ttgcgctcac tgcccgcttt
     3901 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3961 cggtttgcgt attgggcg
//
