LOCUS       sequence-263892-        4125 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 4125)
  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 4125)
  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_SLC32A1
FEATURES             Location/Qualifiers
     source          1..4125
                     /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(2250..2349)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2366..2385)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2367..2385)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2390..2406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2390..2406)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2508..2527)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2519..3328
                     /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(2586..2605)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3475..4063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3964..3983
                     /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 cactgctgag gtctaagctg agcaacgtgg ccaccagcgt
      721 gtccaataag tctcaggcca agatgagcgg catgttcgcc cgcatgggct ttcaggcagc
      781 aacagacgag gaggcagtgg gattcgcaca ctgcgacgat ctggactttg agcaccggca
      841 gggcctgcag atggatatcc tgaaggccga gggcgagcca tgtggcgacg agggagcaga
      901 ggcaccagtg gagggcgata tccactacca gagaggatcc ggagcaccac tgccaccttc
      961 tggcagcaag gaccaagtgg gaggaggagg agagttcgga ggccacgata agcctaagat
     1021 cacagcctgg gaggccggct ggaacgtgac caatgccatc cagggcatgt ttgtgctggg
     1081 actgccatac gcaatcctgc acggaggcta tctgggcctg ttcctgatca tctttgccgc
     1141 cgtggtgtgc tgttacaccg gcaagatcct gatcgcctgc ctgtatgagg agaacgagga
     1201 cggcgaggtg gtgcgggtga gagatagcta tgtggccatc gccaacgcct gctgtgcacc
     1261 taggttccca acactgggag gaagggtggt gaatgtggca cagatcatcg agctggtcat
     1321 gacctgcatc ctgtacgtgg tggtgagcgg caacctgatg tataattcct tcccaggact
     1381 gcccgtgagc cagaagtcct ggtctatcat cgccacagcc gtgctgctgc cctgtgcctt
     1441 tctgaagaac ctgaaggccg tgtccaagtt ctctctgctg tgcaccctgg cccactttgt
     1501 gatcaatatc ctggtcatcg cctactgtct gtctagggca agggactggg catgggagaa
     1561 ggtgaagttc tatatcgatg tgaagaagtt tcctatcagc atcggcatca tcgtgttcag
     1621 ctacacatcc cagatctttc tgccttctct ggagggcaac atgcagcagc caagcgagtt
     1681 ccactgcatg atgaattgga cccacatcgc agcatgcgtg ctgaagggac tgtttgcact
     1741 ggtggcatac ctgacctggg cagacgagac aaaggaagtg atcaccgata acctgcctgg
     1801 cagcatcagg gccgtggtga atatcttcct ggtggccaag gccctgctgt cctatccact
     1861 gcccttcttt gccgccgtgg aggtgctgga gaagtccctg ttccaggagg gctctcgcgc
     1921 cttctttcca gcatgctaca gcggcgatgg ccggctgaag tcctggggac tgacactgag
     1981 atgtgccctg gtggtgttca ccctgctgat ggccatctat gtgccacact ttgcactgct
     2041 gatgggactg accggatccc tgacaggagc aggactgtgc ttcctgctgc cctctctgtt
     2101 tcacctgcgg ctgctgtgga gaaagctgct gtggcaccag gtgttctttg acgtggccat
     2161 cttcgtgatc ggcggcatct gtagcgtgtc cggctttgtg cactccctgg agggcctgat
     2221 cgaggcctac cggaccaatg ccgaggatga cccagctttc ttgtacaaag ttggcattat
     2281 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2341 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2401 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2461 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2521 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2581 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2641 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2701 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2761 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2821 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2881 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2941 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3001 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3061 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3121 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3181 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3241 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3301 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3361 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3421 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3601 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
