LOCUS       sequence-259224-        3945 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 3945)
  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 3945)
  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_SLC16A3
FEATURES             Location/Qualifiers
     source          1..3945
                     /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(2070..2169)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2186..2205)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2187..2205)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2210..2226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2210..2226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2328..2347)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2339..3148
                     /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(2406..2425)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3295..3883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3784..3803
                     /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 accatgggag gagcagtggt ggacgaggga cctaccggag tgaaggcacc
      721 agatggagga tggggatggg ccgtgctgtt cggatgcttt gtgatcacag gcttcagcta
      781 cgcctttccc aaggccgtgt ccgtgttctt taaggagctg atccaggagt tcggcatcgg
      841 ctactccgac accgcctgga tcagctccat cctgctggcc atgctgtatg gcacaggccc
      901 tctgtgcagc gtgtgcgtga acaggtttgg atgcaggcca gtgatgctgg tgggaggact
      961 gttcgcaagc ctgggaatgg tggcagcatc tttttgtagg agcatcatcc aggtgtacct
     1021 gaccacaggc gtgatcaccg gactgggact ggccctgaac ttccagccat ccctgatcat
     1081 gctgaatcgc tatttttcta agcggagacc catggcaaac ggactggcag cagcaggatc
     1141 tcccgtgttc ctgtgcgcac tgagccctct gggacagctg ctgcaggacc ggtatggctg
     1201 gagaggcggc tttctgatcc tgggcggcct gctgctgaat tgctgcgtgt gcgccgccct
     1261 gatgcggccc ctggtggtga ccgcacagcc tggatccgga ccacctagac catctaggcg
     1321 cctgctggac ctgagcgtgt tccgggatag aggctttgtg ctgtacgcag tggcagcatc
     1381 cgtgatggtg ctgggcctgt tcgtgccacc cgtgtttgtg gtgtcttatg ccaaggacct
     1441 gggcgtgcct gataccaagg ccgccttcct gctgacaatc ctgggcttca tcgatatctt
     1501 tgcaaggcca gcagcaggat ttgtggcagg actgggcaag gtgcgccctt actccgtgta
     1561 tctgttcagc ttttccatgt tctttaatgg actggcagac ctggcaggaa gcaccgcagg
     1621 cgattacggc ggcctggtgg tgttctgcat cttctttggc atctcctatg gcatggtggg
     1681 cgccctgcag ttcgaggtgc tgatggccat cgtgggcaca cacaagtttt ctagcgccat
     1741 cggactggtg ctgctgatgg aggcagtggc cgtgctggtg ggacctccaa gcggaggcaa
     1801 gctgctggat gcaacccacg tgtacatgta cgtgttcatc ctggcaggag cagaggtgct
     1861 gacatcctct ctgatcctgc tgctgggcaa cttcttttgt atccggaaga agcccaagga
     1921 gccacagcca gaggtggcag cagcagagga ggagaagctg cacaagccac ctgcagactc
     1981 cggagtggat ctgagggagg tggagcactt cctgaaggcc gagccagaga agaatggcga
     2041 ggtggtgcac acccccgaga catctgtgga cccagctttc ttgtacaaag ttggcattat
     2101 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2161 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2221 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2281 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2341 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2401 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2461 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2521 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2581 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2641 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2701 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2761 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2821 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2881 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2941 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3001 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3061 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3121 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3181 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3241 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3421 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3901 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
