LOCUS       sequence-263862-        4620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4620)
  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 4620)
  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_SLC34A2
FEATURES             Location/Qualifiers
     source          1..4620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2745..2844)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2861..2880)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2862..2880)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2885..2901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2885..2901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3003..3022)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3014..3823
                     /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(3081..3100)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3970..4558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4459..4478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggcac catggcctga gctgggcgac gcacagccaa accctgataa
      721 gtacctggag ggagcagcag gacagcagcc aaccgcaccc gacaagagca aggagaccaa
      781 caagacagat aataccgagg cccctgtgac aaagatcgag ctgctgccaa gctattccac
      841 cgccacactg atcgacgagc ccaccgaggt ggacgatcct tggaatctgc caacactgca
      901 ggactctggc atcaagtgga gcgagcggga taccaagggc aagatcctgt gcttcttcca
      961 gggcatcggc agactgatcc tgctgctggg ctttctgtac ttcttcgtgt gctccctgga
     1021 tatcctgagc tccgccttcc agctggtggg aggcaagatg gcaggacagt tcttttccaa
     1081 ctctagcatc atgtctaatc cactgctggg actggtcatc ggcgtgctgg tgaccgtgct
     1141 ggtgcagtcc tctagcacct ctacaagcat cgtggtgagc atggtgtcct ctagcctgct
     1201 gaccgtgagg gccgccatcc ctatcatcat gggcgccaac atcggcacat ccatcaccaa
     1261 tacaatcgtg gccctgatgc aagtgggcga ccgctctgag tttcggagag ccttcgccgg
     1321 agcaaccgtg cacgatttct ttaactggct gtccgtgctg gtgctgctgc ctgtggaggt
     1381 ggccacccac tacctggaga tcatcacaca gctgatcgtg gagtctttcc actttaagaa
     1441 tggcgaggac gcccctgacc tgctgaaagt gatcacaaag cccttcacca agctgatcgt
     1501 gcagctggac aagaaagtga tctctcagat cgccatgaac gatgagaagg ccaagaataa
     1561 gagcctggtg aagatctggt gtaagacctt cacaaacaag acccagatca atgtgacagt
     1621 gccctccacc gccaattgta catccccttc tctgtgctgg acagacggca tccagaactg
     1681 gaccatgaag aatgtgacat ataaggagaa catcgccaag tgccagcaca tctttgtgaa
     1741 tttccacctg cccgatctgg ccgtgggcac catcctgctg atcctgtctc tgctggtgct
     1801 gtgcggctgc ctgatcatga tcgtgaagat cctgggaagc gtgctgaagg gacaggtggc
     1861 aacagtgatc aagaagacaa tcaacaccga cttccctttt ccattcgcct ggctgaccgg
     1921 atacctggca atcctggtgg gagcaggaat gaccttcatc gtgcagtcct ctagcgtgtt
     1981 caccagcgcc ctgacaccac tgatcggaat cggcgtgatc accatcgagc gggcctatcc
     2041 tctgacactg ggcagcaata tcggcaccac aaccacagca atcctggccg ccctggcatc
     2101 ccctggaaac gccctgagat cctctctgca gatcgccctg tgccacttct ttttcaatat
     2161 cagcggcatc ctgctgtggt atcccatccc tttcacccgg ctgccaatca gaatggccaa
     2221 gggcctgggc aacatctccg ccaagtaccg gtggtttgcc gtgttctatc tgatcatctt
     2281 tttctttctg atcccactga ccgtgtttgg cctgtctctg gcaggatggc gggtgctggt
     2341 gggagtgggc gtgcccgtgg tgttcatcat catcctggtg ctgtgcctga gactgctgca
     2401 gagcaggtgc ccacgcgtgc tgcctaagaa gctgcagaac tggaattttc tgccactgtg
     2461 gatgaggagc ctgaagccct gggacgccgt ggtgtccaag tttaccggct gtttccagat
     2521 gcgctgctgt tgctgttgcc gggtgtgctg cagggcatgt tgcctgctgt gcgattgccc
     2581 aaagtgttgc aggtgctcca agtgttgcga ggacctggag gaggcacagg agggacagga
     2641 tgtgccagtg aaggcccccg agaccttcga caacatcaca atctctaggg aggcacaggg
     2701 agaggtgcca gcaagcgatt ccaagacaga gtgcaccgcc ctggacccag ctttcttgta
     2761 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2821 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2881 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2941 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     3001 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     3061 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3121 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3181 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3241 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3301 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3361 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3421 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3481 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3541 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3601 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3661 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3721 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3781 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3841 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3901 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4081 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
