LOCUS       sequence-265200-        3516 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 3516)
  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 3516)
  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_SLC35B1
FEATURES             Location/Qualifiers
     source          1..3516
                     /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(1641..1740)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1757..1776)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1758..1776)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1781..1797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1781..1797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1899..1918)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1910..2719
                     /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(1977..1996)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2866..3454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3355..3374
                     /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 gctcctctag cctggtgcct gaccggctga gactgccact
      721 gtgcttcctg ggcgtgttcg tgtgctactt ctactatggc atcctgcagg agaagatcac
      781 acggggcaag tatggcgagg gcgccaagca ggagacattc acctttgccc tgaccctggt
      841 gtttatccag tgcgtgatca acgccgtgtt cgccaagatc ctgatccagt tctttgacac
      901 agccagagtg gataggaccc gctcttggct gtacgccgcc tgtagcatct cctatctggg
      961 cgccatggtg tcctctaact ccgccctgca gtttgtgaat taccctacac aggtgctggg
     1021 caagtcttgc aagcccatcc ctgtgatgct gctgggcgtg accctgctga agaagaagta
     1081 ccccctggcc aagtacctgt gcgtgctgct gatcgtggca ggagtggccc tgttcatgta
     1141 caagcctaag aaggtggtgg gcatcgagga gcacaccgtg ggctatggcg agctgctgct
     1201 gctgctgagc ctgacactgg acggactgac cggcgtgtcc caggatcaca tgagggccca
     1261 ctaccagaca ggctctaacc acatgatgct gaacatcaat ctgtggagca cactgctgct
     1321 gggcatgggc atcctgttta ccggcgagct gtgggagttc ctgagctttg ccgagcggta
     1381 cccagccatc atctataata tcctgctgtt tggcctgacc tctgccctgg gccagagctt
     1441 catctttatg acagtggtgt atttcggccc cctgacctgc tccatcatca ccacaacccg
     1501 gaagttcttt acaatcctgg cctccgtgat cctgttcgcc aatccaatct ctcccatgca
     1561 gtgggtgggc accgtgctgg tgtttctggg cctgggcctg gatgccaagt tcggcaaggg
     1621 cgccaagaag acatcccacg acccagcttt cttgtacaaa gttggcatta taagaaagca
     1681 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1741 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     1801 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     1861 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     1921 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1981 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2041 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2101 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2161 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2221 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2281 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2341 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2401 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2461 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2521 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2581 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2641 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2701 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     2761 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     2821 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2881 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2941 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3001 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3061 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3121 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3181 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3241 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3301 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3361 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3421 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3481 gttcctggcc ttttgctggc cttttgctca catgtt
//
