LOCUS       sequence-479339-        4623 bp    DNA     circular SYN 31-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 4623)
  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 4623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 31, 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-SLC34A2_STOP
FEATURES             Location/Qualifiers
     source          1..4623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(76..175)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(191..208)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(191..207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(200..222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      271..357
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      449..476
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(635..652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(806..1394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(886..905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1541..2350)
                     /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     2264..2283
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2342..2361
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2463..2479
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2463..2479
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2484..2503
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2484..2502
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2520..2619
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gtcctgctgc tccctccagg tacttatcag ggtttggctg tgcgtcgccc agctcaggcc
       61 atggtgccat ggtggagcct gcttttttgt acaaagttgg cattataaaa aagcattgct
      121 catcaatgtg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggggcc
      181 cgagcttaag actggccgtc gttttacaac gtcgtgactg ggaaaacatc catgctagcg
      241 ttaacgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac
      301 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
      361 ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg
      421 ccataaactg ccaggcatca aactaagcag aaggccatcc tgacggatgg cctttttgcg
      481 tttctacaaa ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg
      541 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      601 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      661 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      721 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      781 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      841 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      901 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      961 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1021 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1081 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1141 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1201 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1261 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1321 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1381 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1441 acgcgtaact cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg
     1501 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     1561 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1621 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     1681 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     1741 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     1801 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     1861 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     1921 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     1981 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     2041 ctgtttttcc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     2101 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     2161 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     2221 tcccatacaa gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     2281 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa
     2341 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     2401 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct
     2461 gccaggaaac agctatgacc atgtaatacg actcactata ggggatatca gctggatggc
     2521 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat
     2581 gctttcttat aatgccaact ttgtacaaga aagctgggtc ttacagggcg gtgcactctg
     2641 tcttggaatc gcttgctggc acctctccct gtgcctccct agagattgtg atgttgtcga
     2701 aggtctcggg ggccttcact ggcacatcct gtccctcctg tgcctcctcc aggtcctcgc
     2761 aacacttgga gcacctgcaa cactttgggc aatcgcacag caggcaacat gccctgcagc
     2821 acacccggca acagcaacag cagcgcatct ggaaacagcc ggtaaacttg gacaccacgg
     2881 cgtcccaggg cttcaggctc ctcatccaca gtggcagaaa attccagttc tgcagcttct
     2941 taggcagcac gcgtgggcac ctgctctgca gcagtctcag gcacagcacc aggatgatga
     3001 tgaacaccac gggcacgccc actcccacca gcacccgcca tcctgccaga gacaggccaa
     3061 acacggtcag tgggatcaga aagaaaaaga tgatcagata gaacacggca aaccaccggt
     3121 acttggcgga gatgttgccc aggcccttgg ccattctgat tggcagccgg gtgaaaggga
     3181 tgggatacca cagcaggatg ccgctgatat tgaaaaagaa gtggcacagg gcgatctgca
     3241 gagaggatct cagggcgttt ccaggggatg ccagggcggc caggattgct gtggttgtgg
     3301 tgccgatatt gctgcccagt gtcagaggat aggcccgctc gatggtgatc acgccgattc
     3361 cgatcagtgg tgtcagggcg ctggtgaaca cgctagagga ctgcacgatg aaggtcattc
     3421 ctgctcccac caggattgcc aggtatccgg tcagccaggc gaatggaaaa gggaagtcgg
     3481 tgttgattgt cttcttgatc actgttgcca cctgtccctt cagcacgctt cccaggatct
     3541 tcacgatcat gatcaggcag ccgcacagca ccagcagaga caggatcagc aggatggtgc
     3601 ccacggccag atcgggcagg tggaaattca caaagatgtg ctggcacttg gcgatgttct
     3661 ccttatatgt cacattcttc atggtccagt tctggatgcc gtctgtccag cacagagaag
     3721 gggatgtaca attggcggtg gagggcactg tcacattgat ctgggtcttg tttgtgaagg
     3781 tcttacacca gatcttcacc aggctcttat tcttggcctt ctcatcgttc atggcgatct
     3841 gagagatcac tttcttgtcc agctgcacga tcagcttggt gaagggcttt gtgatcactt
     3901 tcagcaggtc aggggcgtcc tcgccattct taaagtggaa agactccacg atcagctgtg
     3961 tgatgatctc caggtagtgg gtggccacct ccacaggcag cagcaccagc acggacagcc
     4021 agttaaagaa atcgtgcacg gttgctccgg cgaaggctct ccgaaactca gagcggtcgc
     4081 ccacttgcat cagggccacg attgtattgg tgatggatgt gccgatgttg gcgcccatga
     4141 tgatagggat ggcggccctc acggtcagca ggctagagga caccatgctc accacgatgc
     4201 ttgtagaggt gctagaggac tgcaccagca cggtcaccag cacgccgatg accagtccca
     4261 gcagtggatt agacatgatg ctagagttgg aaaagaactg tcctgccatc ttgcctccca
     4321 ccagctggaa ggcggagctc aggatatcca gggagcacac gaagaagtac agaaagccca
     4381 gcagcaggat cagtctgccg atgccctgga agaagcacag gatcttgccc ttggtatccc
     4441 gctcgctcca cttgatgcca gagtcctgca gtgttggcag attccaagga tcgtccacct
     4501 cggtgggctc gtcgatcagt gtggcggtgg aatagcttgg cagcagctcg atctttgtca
     4561 caggggcctc ggtattatct gtcttgttgg tctccttgct cttgtcgggt gcggttggct
     4621 gct
//
