LOCUS       sequence-261093-        3573 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 3573)
  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 3573)
  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_SLC25A43
FEATURES             Location/Qualifiers
     source          1..3573
                     /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(1698..1797)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1814..1833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1815..1833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1838..1854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1838..1854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1956..1975)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1967..2776
                     /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(2034..2053)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2923..3511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3412..3431
                     /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 catggcggag agacggcagg ctgaccggag gacagcgcct
      721 gctgtgcgca ggactggccg gcacactgtc tctgagcctg accgcacctc tggagctggc
      781 aacagtgctg gcacaagtgg gagtggtgcg gggacacgcc agaggcccat gggccaccgg
      841 ccacagggtg tggcgcgccg agggactgag ggccctgtgg aagggaaacg cagtggcatg
      901 cctgaggctg ttcccttgtt ccgccgtgca gctggccgcc tacaggaagt tcgtggtgct
      961 gtttacagac gatctgggcc acatctctca gtggagctcc atcatggcag gatctctggc
     1021 aggaatggtg agcaccatcg tgacataccc aaccgacctg atcaagaccc ggctgatcat
     1081 gcagaatatc ctggagccca gctatagagg cctgctgcac gccttctcca caatctacca
     1141 gcaggagggc tttctggccc tgtatagggg cgtgtctctg accgtggtgg gcgccctgcc
     1201 attttccgcc ggatctctgc tggtgtacat gaacctggag aagatctgga atggccctag
     1261 agatcagttc tctctgccac agaactttgc aaacgtgtgc ctggcagcag cagtgaccca
     1321 gacactgagc ttcccctttg agacagtgaa gaggaagatg caggcccagt ccccatatct
     1381 gccacactct ggaggagtgg acgtgcactt ctccggagca gtggattgtt ttcgccagat
     1441 cgtgaaggca cagggcgtgc tgggactgtg gaacggactg acagccaatc tgctgaagat
     1501 cgtgccctac ttcggcatca tgtttagcac cttcgagttt tgcaagagaa tctgtctgta
     1561 ccagaacggc tatatcctga gccccctgtc ctataagctg acccctggag tggatcagtc
     1621 cctgcagcca caggagctga gggagctgaa gaagttcttt aagacaagaa agcccaagcc
     1681 taagaagcct accctggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1741 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1801 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1861 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1921 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1981 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2041 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2101 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2161 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2221 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2281 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2341 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2401 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2461 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2521 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2581 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2641 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2701 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2761 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2821 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3541 cctggccttt tgctggcctt ttgctcacat gtt
//
