LOCUS       sequence-408949-        4338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4338)
  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 4338)
  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-SLC13A1_STOP
FEATURES             Location/Qualifiers
     source          1..4338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      535..544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1895..1994)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2011..2030)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2012..2030)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2035..2051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2035..2051)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2153..2172)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2164..2973
                     /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(2231..2250)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3120..3708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3609..3628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3862..3879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4038..4065)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4157..4243)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4292..4314
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4306..4323
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4307..4323
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgaa gttctttagc
      121 tacatcctgg tgtatcggag attcctgttt gtggtgttta cagtgctggt gctgctgcca
      181 ctgcccatcg tgctgcacac aaaggaggcc gagtgcgcct acaccctgtt tgtggtggcc
      241 acattctggc tgaccgaggc cctgccactg tctgtgaccg ccctgctgcc aagcctgatg
      301 ctgcctatgt tcggcatcat gccttccaag aaggtggcct ctgcctactt taaggacttc
      361 cacctgctgc tgatcggcgt gatctgtctg gccacctcca tcgagaagtg gaacctgcac
      421 aagagaatcg ccctgaagat ggtcatgatg gtgggcgtga atcctgcctg gctgacactg
      481 ggctttatga gctccaccgc cttcctgtct atgtggctgt ctaacaccag cacagccgcc
      541 atggtcatgc caatcgccga ggccgtggtg cagcagatca tcaatgccga ggccgaggtg
      601 gaggccacac agatgaccta cttcaacggc tccaccaatc acggcctgga gatcgacgag
      661 tctgtgaacg gccacgagat caatgagcgg aaggagaaga caaagcctgt gccaggctat
      721 aacaatgata ccggcaagat ctctagcaag gtggagctgg agaagaacag cggcatgagg
      781 acaaagtacc gcaccaagaa gggccacgtg acacggaagc tgacctgcct gtgcatcgcc
      841 tattcctcta ccatcggcgg cctgaccaca atcaccggca catccaccaa cctgatcttt
      901 gccgagtact tcaatacacg gtatcctgac tgcagatgtc tgaactttgg cagctggttc
      961 accttttcct tcccagccgc cctgatcatc ctgctgctgt cctggatctg gctgcagtgg
     1021 ctgtttctgg gctttaattt caaggagatg ttcaagtgcg gcaagacaaa gaccgtgcag
     1081 cagaaggcct gtgccgaagt gatcaagcag gagtaccaga agctgggccc catcaggtat
     1141 caggagatcg tgaccctggt gctgttcatc atcatggccc tgctgtggtt ttctagggac
     1201 ccaggattcg tgcctggatg gagcgccctg ttttccgagt accctggctt cgccacagat
     1261 agcaccgtgg ccctgctgat cggcctgctg ttctttctga tccccgccaa gacactgacc
     1321 aagaccacac ctacaggcga gatcgtggcc tttgattata gcccactgat cacctggaag
     1381 gagtttcagt ccttcatgcc atgggacatc gcaatcctgg tgggaggagg attcgcactg
     1441 gcagatggat gcgaggagtc cggactgtct aagtggatcg gcaataagct gtctccactg
     1501 ggaagcctgc cagcatggct gatcatcctg atcagctccc tgatggtgac atccctgacc
     1561 gaggtggcct ctaaccccgc cacaatcacc ctgttcctgc caatcctgag cccactggca
     1621 gaggcaatcc acgtgaatcc tctgtacatc ctgatcccat ctacactgtg caccagcttt
     1681 gccttcctgc tgccagtggc aaacccacct aatgccatcg tgttttccta tggccacctg
     1741 aaagtgatcg acatggtgaa ggcaggactg ggagtgaaca tcgtgggagt ggccgtggtc
     1801 atgctgggca tctgtacatg gatcgtgcca atgttcgatc tgtacaccta tcctagctgg
     1861 gccccagcca tgtccaatga gaccatgccc taagacccag ctttcttgta caaagttggc
     1921 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1981 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     2041 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     2101 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     2161 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     2221 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2281 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     2341 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     2401 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     2461 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     2521 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     2581 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2641 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2701 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2761 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2821 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2881 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2941 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     3001 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     3061 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3121 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3181 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3241 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3301 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3361 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3421 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3481 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3541 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3601 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3661 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3721 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3781 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3841 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3901 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3961 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     4021 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     4081 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     4141 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     4201 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     4261 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     4321 agtcttaagc tcgggccc
//
