LOCUS       sequence-334675-        3414 bp    DNA     circular SYN 14-APR-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 3414)
  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 3414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-SLC25A10_STOP
FEATURES             Location/Qualifiers
     source          1..3414
                     /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(971..1070)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1087..1106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1088..1106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1111..1127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1111..1127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1229..1248)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1240..2049
                     /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(1307..1326)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2196..2784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2685..2704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2938..2955
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3114..3141)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3233..3319)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3368..3390
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3382..3399
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3383..3399
                     /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 ccaccatggc agcagaggca
      121 agggtgtcta ggtggtactt cggaggactg gcaagctgcg gagcagcatg ctgtacacac
      181 ccactggacc tgctgaaggt gcacctgcag acccagcagg aggtgaagct gaggatgaca
      241 ggcatggccc tgcgggtggt gagaaccgat ggcatcctgg ccctgtattc cggactgtct
      301 gccagcctgt gcaggcagat gacctactct ctgacaagat ttgccatcta tgagacagtg
      361 cgggacagag tggcaaaggg atcccaggga ccactgcctt tccacgagaa ggtgctgctg
      421 ggatccgtgt ctggactggc aggaggattt gtgggaaccc ctgcagacct ggtgaacgtg
      481 cggatgcaga atgatgtgaa gctgccacag ggccagcgga gaaactacgc ccacgccctg
      541 gatggactgt atagggtggc aagggaggag ggactgaggc gcctgttcag cggagcaaca
      601 atggcaagct ccagaggcgc cctggtgacc gtgggacagc tgtcctgcta cgaccaggcc
      661 aagcagctgg tgctgagcac aggctatctg tccgataaca tcttcaccca ctttgtggca
      721 tctttcatcg caggaggatg cgccaccttc ctgtgccagc cactggacgt gctgaagacc
      781 agactgatga atagcaaggg cgagtaccag ggcgtgttcc actgtgcagt ggagaccgca
      841 aagctgggac ctctggcctt ttataagggc ctggtgccag ccggcatcag gctgatcccc
      901 cacaccgtgc tgacattcgt gtttctggag cagctgcgca agaattttgg catcaaggtg
      961 ccctcttaag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1021 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1081 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     1141 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1201 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1261 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1321 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     1381 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1441 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     1501 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     1561 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     1621 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     1681 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     1741 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     1801 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     1861 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     1921 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     1981 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2041 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     2101 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     2161 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2221 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2281 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2341 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2401 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2461 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2521 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2581 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2641 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2701 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2761 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2821 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2881 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2941 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3001 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3061 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     3121 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     3181 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     3241 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     3301 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     3361 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccc
//
