LOCUS       sequence-262701-        4260 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 4260)
  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 4260)
  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_SLC47A1
FEATURES             Location/Qualifiers
     source          1..4260
                     /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(2385..2484)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2501..2520)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2502..2520)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2525..2541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2525..2541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2643..2662)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2654..3463
                     /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(2721..2740)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3610..4198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4099..4118
                     /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 accatggagg cccctgagga gccagcacct gtgagaggag gaccagaggc
      721 caccctggag gtgaggggct cccgctgcct gcggctgtct gccttcagag aggagctgag
      781 ggccctgctg gtgctggcag gaccagcctt tctggtgcag ctgatggtgt tcctgatctc
      841 ttttatcagc tccgtgttct gcggacacct gggcaagctg gagctggacg cagtgaccct
      901 ggcaatcgcc gtgatcaacg tgacaggcgt gagcgtgggc tttggcctgt ctagcgcctg
      961 tgataccctg atcagccaga catacggctc ccagaatctg aagcacgtgg gcgtgatcct
     1021 gcagaggtcc gccctggtgc tgctgctgtg ctgtttccca tgttgggccc tgtttctgaa
     1081 cacccagcac atcctgctgc tgttcagaca ggaccccgac gtgagcaggc tgacccagac
     1141 atacgtgacc atcttcatcc ctgccctgcc agccacattt ctgtacatgc tgcaggtgaa
     1201 gtatctgctg aatcagggca tcgtgctgcc acagatcgtg acaggagtgg cagccaacct
     1261 ggtgaatgcc ctggccaact atctgttcct gcaccagctg cacctgggcg tgatcggaag
     1321 cgccctggcc aacttgatct ctcaatacac gcttgcactg ctgctgtttc tgtatatcct
     1381 gggcaagaag ctgcaccagg caacatgggg aggatggagc ctggagtgcc tgcaggactg
     1441 ggcatccttc ctgaggctgg caatcccctc tatgctgatg ctgtgcatgg agtggtgggc
     1501 ctatgaagtg ggcagctttc tgtccggaat cctgggaatg gtggagctgg gagcacagag
     1561 catcgtgtac gagctggcca tcatcgtgta tatggtgcct gccggcttct ccgtggccgc
     1621 ctctgtgcgc gtgggaaacg cactgggagc aggcgacatg gagcaggccc ggaagtcctc
     1681 taccgtgtcc ctgctgatca cagtgctgtt cgccgtggcc ttttctgtgc tgctgctgag
     1741 ctgcaaggat cacgtgggct acatcttcac cacagaccgc gatatcatca atctggtggc
     1801 ccaggtggtg cctatctatg ccgtgagcca cctgtttgag gccctggcct gtacctctgg
     1861 aggcgtgctg aggggaagcg gaaaccagaa agtgggagcc atcgtgaata caatcggcta
     1921 ctatgtggtg ggcctgccaa tcggcatcgc cctgatgttc gccaccacac tgggcgtgat
     1981 gggcctgtgg tccggcatca tcatctgcac cgtgtttcag gccgtgtgct tcctgggctt
     2041 tatcatccag ctgaactgga agaaggcatg ccagcaggca caggtgcacg ccaacctgaa
     2101 ggtgaacaat gtgcctcggt ctggcaatag cgccctgcca caggaccctc tgcacccagg
     2161 atgtcccgag aacctggagg gcatcctgac caatgacgtg ggcaagacag gcgagcccca
     2221 gagcgatcag cagatgagac aggaggagcc actgccagag caccctcagg atggagccaa
     2281 gctgtccagg aagcagctgg tgctgaggag aggactgctg ctgctgggcg tgttcctgat
     2341 cctgctggtg ggcatcctgg tgcgctttta cgtgcggatc caggacccag ctttcttgta
     2401 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2461 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2521 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2581 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2641 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2701 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2761 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2821 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2881 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2941 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3001 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3061 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3121 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3181 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3241 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3301 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3361 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3421 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3481 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3541 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3601 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3661 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3721 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3901 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4141 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4201 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
