LOCUS       sequence-262932-        3561 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 3561)
  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 3561)
  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_SLC35A1
FEATURES             Location/Qualifiers
     source          1..3561
                     /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(1686..1785)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1802..1821)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1803..1821)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1826..1842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1826..1842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1944..1963)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1955..2764
                     /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(2022..2041)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2911..3499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3400..3419
                     /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 accatggcag cacctaggga caacgtgacc ctgctgttca agctgtactg
      721 cctggccgtg atgaccctga tggcagccgt gtacacaatc gccctgcggt ataccagaac
      781 atctgataag gagctgtatt tcagcaccac agccgtgtgc atcaccgaag tgatcaagct
      841 gctgctgagc gtgggcatcc tggccaagga gacaggctcc ctgggcaggt ttaaggcctc
      901 tctgcgcgag aatgtgctgg gctcccccaa ggagctgctg aagctgtctg tgcctagcct
      961 ggtgtacgcc gtgcagaaca atatggcctt tctggccctg agcaacctgg acgcagccgt
     1021 gtaccaggtg acctatcagc tgaagatccc atgcaccgcc ctgtgcacag tgctgatgct
     1081 gaataggaca ctgagcaagc tgcagtgggt gtccgtgttc atgctgtgcg caggagtgac
     1141 cctggtgcag tggaagccag cccaggccac aaaggtggtg gtggagcaga accccctgct
     1201 gggctttgga gcaatcgcaa tcgccgtgct gtgctccgga ttcgcaggcg tgtactttga
     1261 gaaggtgctg aagagctccg acacctccct gtgggtgcgc aatatccaga tgtacctgtc
     1321 tggcatcatc gtgacactgg ccggcgtgta tctgtctgat ggcgccgaga tcaaggagaa
     1381 gggcttcttt tacggctata cctactacgt gtggttcgtg atctttctgg caagcgtggg
     1441 aggactgtac acctccgtgg tggtgaagta tacagacaac atcatgaagg gcttctctgc
     1501 cgcagcagca atcgtgctgt ccaccatcgc ctctgtgatg ctgttcggcc tgcagatcac
     1561 cctgacattt gccctgggca cactgctggt gtgcgtgagc atctacctgt atggcctgcc
     1621 ccggcaggat accacatcta tccagcaggg cgagaccgcc agcaaggaga gagtgatcgg
     1681 cgtggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1741 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1801 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1861 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1921 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1981 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2041 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2101 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2161 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2221 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2281 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2341 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2401 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2461 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2521 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2581 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2641 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2701 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2761 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2821 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2881 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2941 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3001 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3061 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3121 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3181 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3241 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3301 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3361 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3421 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3481 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3541 ctggcctttt gctcacatgt t
//
