LOCUS       sequence-260408-        4176 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 4176)
  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 4176)
  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_SLC22A8
FEATURES             Location/Qualifiers
     source          1..4176
                     /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(2301..2400)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2417..2436)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2418..2436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2441..2457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2441..2457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2559..2578)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2570..3379
                     /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(2637..2656)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3526..4114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4015..4034
                     /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 accatgacct tctctgagat cctggaccgg gtgggcagca tgggccactt
      721 ccagtttctg cacgtggcca tcctgggcct gcctatcctg aacatggcca accacaatct
      781 gctgcagatc ttcaccgcag caacaccagt gcaccactgc aggccacctc acaacgcatc
      841 taccggacca tgggtgctgc caatgggacc taatggcaag cctgagcggt gtctgagatt
      901 tgtgcaccca cccaacgcca gcctgcctaa tgacacacag cgcgccatgg agccatgcct
      961 ggatggctgg gtgtacaata gcaccaagga ctccatcgtg acagagtggg acctggtgtg
     1021 caactccaat aagctgaagg agatggccca gtctatcttc atggcaggaa tcctgatcgg
     1081 aggactggtg ctgggcgacc tgtctgatcg gtttggccgg agacctatcc tgacctgcag
     1141 ctacctgctg ctggcagcaa gcggatccgg agcagccttc tccccaacat ttcccatcta
     1201 tatggtgttc cggttcctgt gcggcttcgg catctctggc atcaccctga gcacagtgat
     1261 cctgaacgtg gagtgggtgc caaccaggat gcgcgccatc atgtccaccg ccctgggcta
     1321 ctgctataca ttcggccagt ttatcctgcc aggactggca tacgcaatcc cacagtggag
     1381 gtggctgcag ctgaccgtgt ctatcccctt cttcgtgttc ttcctgagct cctggtggac
     1441 acccgagtcc atccgctggc tggtgctgtc tggcaagtct agcaaggccc tgaagatcct
     1501 gaggcgcgtg gccgtgttca acggcaagaa ggaggaggga gagcggctga gcctggagga
     1561 gctgaagctg aatctgcaga aggagatcag cctggccaag gccaagtata ccgcctccga
     1621 cctgttcaga atccctatgc tgcggagaat gaccttttgt ctgtccctgg cctggttcgc
     1681 cacaggcttt gcctactatt ctctggccat gggcgtggag gagtttggcg tgaatctgta
     1741 catcctgcag atcatcttcg gcggcgtgga cgtgccagcc aagtttatca ccatcctgtc
     1801 tctgagctat ctgggcaggc acaccacaca ggcagccgcc ctgctgctgg caggaggagc
     1861 catcctggcc ctgacattcg tgcccctgga tctgcagacc gtgcgcacag tgctggccgt
     1921 gtttggcaag ggctgcctgt cctctagctt cagctgtctg tttctgtaca cctccgagct
     1981 gtatcccacc gtgatcaggc agacaggcat gggcgtgtcc aacctgtgga ccagggtggg
     2041 ctccatggtg tctcctctgg tgaagatcac aggcgaggtg cagcctttca tcccaaatat
     2101 catctatggc atcacagccc tgctgggagg aagcgccgcc ctgtttctgc ccgagaccct
     2161 gaaccagcct ctgccagaga caatcgagga tctggagaat tggagcctgc gggccaagaa
     2221 gcccaagcag gagcctgagg tggagaaggc ctcccagaga atcccactgc agcctcacgg
     2281 accaggactg ggctcctctg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2341 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2401 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2461 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2521 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2581 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2641 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2701 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2761 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2821 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2881 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2941 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3001 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3061 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3121 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3181 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3241 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3301 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3361 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3421 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3481 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3541 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3601 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3661 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3721 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3781 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3841 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3901 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3961 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4021 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4081 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4141 gttcctggcc ttttgctggc cttttgctca catgtt
//
