LOCUS       sequence-262895-        4356 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 4356)
  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 4356)
  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_SLC47A2
FEATURES             Location/Qualifiers
     source          1..4356
                     /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(2481..2580)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2597..2616)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2598..2616)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2621..2637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2621..2637)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2739..2758)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2750..3559
                     /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(2817..2836)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3706..4294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4195..4214
                     /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 accatggact ccctgcagga taccgtggcc ctggaccacg gaggatgctg
      721 tcccgccctg tctaggctgg tgcctagagg attcggcacc gagatgtgga cactgtttgc
      781 cctgtccggc cccctgttcc tgtttcaggt gctgacattc atgatctaca tcgtgtccac
      841 cgtgttttgc ggacacctgg gcaaggtgga gctggcatct gtgaccctgg cagtggcctt
      901 cgtgaacgtg tgcggcgtga gcgtgggagt gggactgagc tccgcctgtg acacactgat
      961 gtctcagagc tttggctccc cgaataaaaa acatgtagga gtcatcctgc agagaggcgc
     1021 cctggtgctg ctgctgtgct gtctgccctg ttgggccctg ttcctgaaca cccagcacat
     1081 cctgctgctg tttaggcagg accctgatgt gagccgcctg acacaggatt acgtgatgat
     1141 cttcatccca ggcctgcccg tgatctttct gtacaacctg ctggccaagt atctgcagaa
     1201 tcagggctgg ctgaagggcc aggaggagga gagccctttc cagaccccag gcctgtccat
     1261 cctgcaccca tcccactctc acctgtctag ggccagcttc cacctgtttc agaagatcac
     1321 ctggccacag gtgctgagcg gagtggtggg caactgcgtg aatggcgtgg ccaactacgc
     1381 cctggtgtcc gtgctgaatc tgggcgtgcg cggctctgcc tatgccaata tcatcagcca
     1441 gttcgcccag accgtgtttc tgctgctgta catcgtgctg aagaagctgc acctggagac
     1501 atgggcagga tggtctagcc agtgcctgca ggactggggc ccattctttt ctctggccgt
     1561 gcccagcatg ctgatgatct gcgtggagtg gtgggcctat gagatcggct ccttcctgat
     1621 gggcctgctg tccgtggtgg atctgtctgc ccaggccgtg atctacgagg tggccaccgt
     1681 gacatatatg atcccactgg gactgagcat cggcgtgtgc gtgcgggtgg gaatggcact
     1741 gggagcagcc gataccgtgc aggcaaagcg gagcgccgtg agcggcgtgc tgagcatcgt
     1801 gggcatctcc ctggtgctgg gcaccctgat cagcatcctg aagaaccagc tgggccacat
     1861 ctttacaaat gacgaggatg tgatcgccct ggtgtcccag gtgctgcccg tgtacagcgt
     1921 gttccacgtg tttgaggcca tctgctgcgt gtacggaggc gtgctgaggg gaacaggcaa
     1981 gcaggccttc ggcgcagcag tgaacgcaat cacctactat atcatcggcc tgccactggg
     2041 catcctgctg acatttgtgg tgaggatgcg catcatggga ctgtggctgg gaatgctggc
     2101 atgcgtgttc ctggcaacag cagcctttgt ggcatacacc gcaaggctgg actggaagct
     2161 ggcagcagag gaggccaaga agcactctgg caggcagcag cagcagaggg cagagagcac
     2221 cgcaacaaga ccaggacctg agaaggccgt gctgtcctct gtggccacag gcagctcccc
     2281 tggcatcacc ctgaccacat atagccggtc cgagtgtcac gtggatttct ttagaacacc
     2341 tgaggaggca cacgcactgt ctgccccaac cagcaggctg tccgtgaagc agctggtcat
     2401 ccggagagga gccgccctgg gagcagcaag cgccaccctg atggtgggcc tgacagtgcg
     2461 gatcctggcc accagacacg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2521 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2581 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2641 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2701 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2761 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2821 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2881 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2941 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3001 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3061 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3121 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3181 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3241 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3301 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3361 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3421 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3481 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3541 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3601 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3661 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3721 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3781 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3841 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3901 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3961 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4021 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4081 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4141 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4201 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4261 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4321 gttcctggcc ttttgctggc cttttgctca catgtt
//
