LOCUS       sequence-458141-        5907 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 5907)
  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 5907)
  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-SLC4A10_STOP
FEATURES             Location/Qualifiers
     source          1..5907
                     /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(4032..4131)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4148..4167)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4149..4167)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4172..4188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4172..4188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4290..4309)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4301..5110
                     /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(4368..4387)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5257..5845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5746..5765
                     /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 accatggaga tcaaggatca gggcgcccag atggagcctc tgctgccaac
      721 ccggaacgat gaggaggccg tggtggacag gggaggaacc agatccatcc tgaagacaca
      781 cttcgagaag gaggacctgg agggccaccg gacactgttt atcggagtgc acgtgccact
      841 gggaggaaga aagagccacc ggagacacag gcacagggga cacaagcacc ggaagagaga
      901 tagggagagg gactctggac tggaggatgg aagggagagc ccttcctttg acaccccaag
      961 ccagagagtg cagttcatcc tgggcacaga ggacgatgac gaggagcaca tcccccacga
     1021 tctgttcacc gagctggacg agatctgctg gagagaagga gaggacgcag agtggaggga
     1081 gacagcaagg tggctgaagt ttgaggagga tgtggaggac ggaggagagc ggtggtccaa
     1141 gccttacgtg gccaccctgt ccctgcactc tctgttcgag ctgagatctt gtatcctgaa
     1201 cggcacagtg ctgctggata tgcacgccaa taccctggag gagatcgccg atatggtgct
     1261 ggaccagcag gtgagctccg gccagctgaa tgaggacgtg cggcacagag tgcacgaggc
     1321 cctgatgaag cagcaccacc accagaacca gaagaagctg accaatcgga tcccaatcgt
     1381 gagatccttc gccgatatcg gcaagaagca gtctgagccc aacagcatgg acaagaatgc
     1441 aggacaggtg gtgagcccac agtccgcccc tgcctgcgtg gagaacaaga atgacgtgag
     1501 cagggagaac tctaccgtgg actttagcaa gggactggga ggacagcaga agggacacac
     1561 atccccctgt ggcatgaagc agaggcacga gaagggacca cctcaccagc aggagaggga
     1621 ggtggatctg cacttcatga agaagatccc acccggcgcc gaggcctcca atatcctggt
     1681 gggcgagctg gagtttctgg accggaccgt ggtggccttc gtgagactgt ctccagccgt
     1741 gctgctgcag ggactggcag aggtgccaat cccaacacgg ttcctgttta tcctgctggg
     1801 acctctgggc aagggacagc agtaccacga gatcggcaga tccatcgcca ccctgatgac
     1861 agatgaggtg ttccacgacg tggcctataa ggccaaggat aggaacgacc tggtgagcgg
     1921 catcgatgag tttctggacc aggtgaccgt gctgcctcca ggcgagtggg atccctctat
     1981 caggatcgag ccccctaaga acgtgcctag ccaggagaag cgcaagatcc ccgccgtgcc
     2041 taatggaacc gcagcacacg gagaggcaga gccacacggc ggccacagcg gacctgagct
     2101 gcagaggaca ggacgcatct ttggcggcct gatcctggac atcaagcgga aggccccata
     2161 cttctggagc gacttccggg acgccttctc cctgcagtgc ctggcctctt tcctgtttct
     2221 gtattgcgcc tgtatgtccc ccgtgatcac attcggcgga ctgctgggag aggcaaccga
     2281 gggaaggatc agcgccatcg agtccctgtt cggcgcctct atgaccggca tcgcctacag
     2341 cctgtttgga ggacagccac tgaccatcct gggatccaca ggacccgtgc tggtgttcga
     2401 gaagatcctg ttcaagtttt gcaaggagta cggcctgtct tatctgagcc tgagggcaag
     2461 catcggactg tggaccgcca cactgtgcat catcctggtg gccacagacg cctctagcct
     2521 ggtgtgctac atcacccgct ttacagagga ggccttcgcc agcctgatct gtatcatctt
     2581 tatctacgag gccctggaga agctgttcga gctgtccgag gcctatccaa tcaacatgca
     2641 caatgatctg gagctgctga cccagtattc ttgcaactgc gtggagccac acaaccccag
     2701 caatggcaca ctgaaggagt ggcgggagtc caacatctct gccagcgaca tcatctggga
     2761 gaatctgacc gtgtctgagt gcaagagcct gcacggcgag tacgtgggca gagcctgtgg
     2821 ccacgatcac ccttatgtgc cagacgtgct gttctggtct gtgatcctgt tctttagcac
     2881 cgtgacactg tccgccaccc tgaagcagtt taagacaagc aggtacttcc ccaccaaggt
     2941 gcgctccatc gtgtctgatt tcgccgtgtt tctgaccatc ctgtgcatgg tgctgatcga
     3001 ctatgccatc ggcatccctt ccccaaagct gcaggtgccc agcgtgttca agcctacacg
     3061 ggatgacaga ggctggtttg tgacccctct gggccccaac ccttggtgga cagtgatcgc
     3121 cgccatcatc ccagccctgc tgtgcaccat cctgatcttt atggatcagc agatcacagc
     3181 cgtgatcatc aatagaaagg agcacaagct gaagaagggc tgcggctatc acctggacct
     3241 gctgatggtg gccgtgatgc tgggcgtgtg ctccatcatg ggactgcctt ggttcgtggc
     3301 agcaaccgtg ctgagcatca cacacgtgaa ctccctgaag ctggagagcg agtgctccgc
     3361 cccaggagag cagccaaagt tcctgggaat cagggagcag agggtgaccg gcctgatgat
     3421 ctttatcctg atgggctcca gcgtgttcat gaccagcatc ctgaagttca tcccaatgcc
     3481 cgtgctgtac ggcgtgtttc tgtatatggg cgccagctcc ctgaagggca tccagttctt
     3541 tgataggatc aagctgttct ggatgccagc caagcaccag cccgacttta tctacctgag
     3601 gcacgtgccc ctgcgcaagg tgcacctgtt caccatcatc cagatgtcct gtctgggcct
     3661 gctgtggatc atcaaggtgt ctcgcgccgc catcgtgttt cctatgatgg tgctggccct
     3721 ggtgttcgtg aggaagctga tggatctgct gtttaccaag cgcgagctga gctggctgga
     3781 tgacctgatg ccagagagca agaagaagaa gctggaggat gccgagaagg aggaggagca
     3841 gtctatgctg gccatggagg acgagggcac agtgcagctg cctctggagg gccactatag
     3901 ggatgaccca agcgtgatca atatctccga cgagatgtct aagaccgccc tgtggcgcaa
     3961 cctgctgatc acagccgata atagcaagga caaggagtct agcttccctt ctaagtcctc
     4021 tccaagctaa gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     4081 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     4141 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     4201 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     4261 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     4321 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     4381 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     4441 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     4501 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     4561 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     4621 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     4681 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4741 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4801 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4861 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4921 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4981 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     5041 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     5101 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     5161 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     5221 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5281 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5341 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5401 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5461 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5521 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5581 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5641 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5701 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5761 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5821 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5881 cttttgctgg ccttttgctc acatgtt
//
