LOCUS       sequence-312310-        4509 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 4509)
  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 4509)
  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-SLC20A2_STOP
FEATURES             Location/Qualifiers
     source          1..4509
                     /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(2634..2733)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2750..2769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2751..2769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2774..2790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2774..2790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2892..2911)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2903..3712
                     /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(2970..2989)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3859..4447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4348..4367
                     /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 accatggcca tggacgagta tctgtggatg gtcatcctgg gcttcatcat
      721 cgcctttatc ctggccttca gcgtgggcgc caacgatgtg gccaattcct ttggcaccgc
      781 agtgggatct ggagtggtga cactgaggca ggcctgcatc ctggccagca tcttcgagac
      841 cacaggatcc gtgctgctgg gagcaaaagt gggagagacc atccgcaagg gcatcatcga
      901 cgtgaacctg tacaatgaga ccgtggagac actgatggca ggagaggtga gcgccatggt
      961 gggatccgcc gtgtggcagc tgatcgcatc ttttctgcgg ctgcccatca gcggcaccca
     1021 ctgtatcgtg ggctctacaa tcggcttcag cctggtggcc atcggcacaa agggcgtgca
     1081 gtggatggag ctggtgaaga tcgtggcctc ttggttcatc agccctctgc tgtccggctt
     1141 catgtctggc ctgctgtttg tgctgatccg gatcttcatc ctgaagaagg aggacccagt
     1201 gcctaacgga ctgagagccc tgccagtgtt ttatgccgcc accatcgcca tcaacgtgtt
     1261 cagcatcatg tacacaggag cacctgtgct gggactggtg ctgccaatgt gggccatcgc
     1321 cctgatctct tttggcgtgg ccctgctgtt cgccttcttc gtgtggctgt tcgtgtgccc
     1381 ctggatgcgg agaaagatca ccggcaagct gcagaaggag ggcgccctgt ccagggtgtc
     1441 tgatgagagc ctgtccaagg tgcaggaggc cgagtctcca gtgttcaagg agctgccagg
     1501 agcaaaggca aacgacgata gcaccatccc tctgacagga gcagcaggag agaccctggg
     1561 cacatccgag ggcaccagcg ccggatccca ccctagggca gcatacggaa gagccctgag
     1621 catgacacac ggctctgtga agagcccaat ctccaatggc accttcggct ttgacggcca
     1681 cacacgcagc gatggccacg tgtatcacac cgtgcacaag gactccggcc tgtacaagga
     1741 tctgctgcac aagatccaca tcgacagggg accagaggag aagcctgcac aggagtccaa
     1801 ctatcgcctg ctgaggcgca acaattctta cacctgctat acagcagcaa tctgtggact
     1861 gccagtgcac gccaccttcc gggcagcaga cagctccgcc ccagaggatt ctgagaagct
     1921 ggtgggcgac acagtgtctt acagcaagaa gaggctgcgc tacgatagct attctagcta
     1981 ctgcaatgca gtggcagagg cagagatcga ggcagaggag ggcggcgtgg agatgaagct
     2041 ggcatccgag ctggcagacc cagatcagcc aagagaggac ccagcagagg aggagaagga
     2101 ggagaaggat gcccctgagg tgcacctgct gttccacttt ctgcaggtgc tgaccgcatg
     2161 cttcggaagc tttgcccacg gaggcaacga cgtgagcaat gcaatcggac cactggtggc
     2221 cctgtggctg atctataagc agggaggagt cacccaggag gcagcaacac ccgtgtggct
     2281 gctgttttac ggaggagtgg gaatctgtac cggactgtgg gtgtggggcc ggagagtgat
     2341 ccagacaatg ggcaaggacc tgacccctat cacaccatcc tctggcttta ccatcgagct
     2401 ggccagcgcc ttcacagtgg tcatcgcctc caacatcggc ctgcccgtgt ctaccacaca
     2461 ctgtaaagtg ggatctgtgg tggcagtggg atggatccgg agcagaaagg ccgtggattg
     2521 gaggctgttt cgcaatatct tcgtggcatg gtttgtgacc gtgccagtgg caggactgtt
     2581 ctccgccgcc gtgatggccc tgctgatgta tggcatcctg ccttacgtgt aagacccagc
     2641 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2701 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2761 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2821 atgttacatt gcacaagata aaataatatc atcatgaaca ataaaactgt ctgcttacat
     2881 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2941 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     3001 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     3061 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     3121 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3181 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3241 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3301 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3361 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3421 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3481 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3541 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3601 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3661 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3721 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3781 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3841 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3901 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3961 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4021 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4081 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4141 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4201 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4261 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4321 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4381 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4441 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4501 tcacatgtt
//
