LOCUS       sequence-338285-        5286 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 5286)
  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 5286)
  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-SLC4A1_STOP
FEATURES             Location/Qualifiers
     source          1..5286
                     /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"
     protein_bind    complement(2843..2942)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2959..2978)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2960..2978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2983..2999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2983..2999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3101..3120)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3112..3921
                     /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(3179..3198)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4068..4656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4557..4576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4810..4827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4986..5013)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5105..5191)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5240..5262
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5254..5271
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5255..5271
                     /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 ccaccatgga ggagctgcag
      121 gacgattacg aggacatgat ggaggagaac ctggagcagg aggagtatga ggaccccgat
      181 atccctgagt cccagatgga ggagccagca gcacacgata ccgaggcaac cgccacagac
      241 taccacacca catctcaccc cggcacacac aaggtgtatg tggagctgca ggagctggtc
      301 atggatgaga agaatcagga gctgcggtgg atggaggccg ccagatgggt gcagctggag
      361 gagaacctgg gagagaatgg agcatggggc agaccccacc tgtcccacct gacattttgg
      421 tctctgctgg agctgcggag agtgttcacc aagggcacag tgctgctgga cctgcaggag
      481 acctctctgg caggagtggc aaaccagctg ctggacaggt ttatcttcga ggatcagatc
      541 aggcctcagg acagggagga gctgctgcgc gccctgctgc tgaagcactc ccacgcagga
      601 gagctggagg ccctgggagg cgtgaagcca gccgtgctga caagatctgg cgatccaagc
      661 cagccactgc tgcctcagca cagctccctg gagacccagc tgttctgcga gcagggcgac
      721 ggaggaacag agggacactc cccatctggc atcctggaga agatcccccc tgatagcgag
      781 gccaccctgg tgctggtggg aagggcagac tttctggagc agcccgtgct gggcttcgtg
      841 agactgcagg aggcagcaga gctggaggca gtggagctgc cagtgcctat ccggtttctg
      901 ttcgtgctgc tgggaccaga ggcaccacac atcgattaca cccagctggg aagagcagca
      961 gccacactga tgagcgagcg ggtgttcaga atcgacgcct atatggcaca gtccagggga
     1021 gagctgctgc actctctgga gggctttctg gattgttccc tggtgctgcc accaaccgac
     1081 gcaccaagcg agcaggccct gctgtccctg gtgccagtgc agagggagct gctgaggcgc
     1141 cggtaccagt ctagccctgc caagccagat tcctctttct ataagggact ggacctgaat
     1201 ggaggaccag acgatcctct gcagcagaca ggccagctgt ttggcggcct ggtgagggac
     1261 atcagaaggc gctaccctta ctatctgagc gacatcaccg atgccttctc cccacaggtg
     1321 ctggcagccg tgatctttat ctatttcgcc gccctgagcc cagcaatcac ctttggcgga
     1381 ctgctgggcg agaagacacg caaccagatg ggcgtgtctg agctgctgat cagcacagcc
     1441 gtgcagggca tcctgttcgc actgctggga gcacagccac tgctggtggt gggcttcagc
     1501 ggccctctgc tggtgtttga ggaggccttc ttttccttct gcgagaccaa tggcctggag
     1561 tacatcgtgg gcagggtgtg gatcggcttc tggctgatcc tgctggtggt gctggtggtg
     1621 gcctttgagg gctctttcct ggtgcggttt atcagcagat acacccagga gatcttttcc
     1681 ttcctgatct ctctgatctt catctatgag accttcagca agctgatcaa gatcttccag
     1741 gatcaccccc tgcagaagac ctacaactat aatgtgctga tggtgccaaa gccacaggga
     1801 cctctgccaa acaccgccct gctgagcctg gtgctgatgg caggcacatt ctttttcgcc
     1861 atgatgctgc gcaagttcaa gaatagctcc tactttcccg gcaagctgcg gagagtgatc
     1921 ggcgatttcg gcgtgcctat cagcatcctg atcatggtgc tggtggactt tttcatccag
     1981 gatacctata cacagaagct gtccgtgcct gacggcttta aggtgtctaa ctctagcgcc
     2041 aggggatggg tcatccaccc tctgggactg cgctccgagt tcccaatctg gatgatgttt
     2101 gcctctgccc tgcctgccct gctggtgttc atcctgatct ttctggagag ccagatcacc
     2161 acactgatcg tgtccaagcc agagcggaag atggtgaagg gctccggctt tcacctggac
     2221 ctgctgctgg tggtgggaat gggaggagtg gccgccctgt tcggaatgcc atggctgtct
     2281 gccaccacag tgagaagcgt gacccacgcc aatgccctga cagtgatggg caaggcatct
     2341 acccctggag cagcagcaca gatccaggag gtgaaggagc agaggatcag cggactgctg
     2401 gtggccgtgc tggtgggact gtctatcctg atggagccta tcctgagccg catcccactg
     2461 gccgtgctgt ttggcatctt cctgtacatg ggcgtgacca gcctgtccgg catccagctg
     2521 tttgatagga tcctgctgct gttcaagcct ccaaagtacc acccagacgt gccctatgtg
     2581 aagagggtga agacctggcg catgcacctg ttcacaggca tccagatcat ctgcctggcc
     2641 gtgctgtggg tggtgaagag caccccagca tccctggccc tgccttttgt gctgatcctg
     2701 acagtgccac tgaggcgcgt gctgctgcca ctgatcttcc ggaacgtgga gctgcagtgt
     2761 ctggatgccg acgatgccaa ggccaccttt gacgaggagg agggcagaga cgagtacgat
     2821 gaggtggcca tgcccgtgta agacccagct ttcttgtaca aagttggcat tataagaaag
     2881 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     2941 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     3001 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     3061 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     3121 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     3181 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     3241 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     3301 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     3361 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     3421 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     3481 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     3541 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     3601 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     3661 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     3721 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     3781 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     3841 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     3901 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     3961 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     4021 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4081 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4141 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4201 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4261 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4321 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4381 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4441 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4501 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4561 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4621 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4681 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4741 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4801 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4861 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4921 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     4981 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     5041 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     5101 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     5161 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
     5221 acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
     5281 gggccc
//
