LOCUS       sequence-369797-        3462 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 3462)
  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 3462)
  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-SLC25A29_STOP
FEATURES             Location/Qualifiers
     source          1..3462
                     /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(1019..1118)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1135..1154)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1136..1154)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1159..1175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1159..1175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1277..1296)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1288..2097
                     /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(1355..1374)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2244..2832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2733..2752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2986..3003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3162..3189)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3281..3367)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3416..3438
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3430..3447
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3431..3447
                     /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 cctggacttc
      121 ctggcaggat gcgcaggagg agtggcaggc gtgctggtgg gccacccttt tgataccgtg
      181 aaggtgcggc tgcaggtgca gtctgtggag aagccacagt acagaggcac cctgcactgt
      241 ttcaagtcca tcatcaagca ggagtctgtg ctgggcctgt ataagggact gggaagccct
      301 ctgatgggac tgacctttat aaatgctctc gtgttcggcg tgcagggcaa tacactgcgg
      361 gccctgggac acgactcccc actgaaccag tttctggcag gagcagcagc aggagcaatc
      421 cagtgcgtga tctgctgtcc aatggagctg gccaagacca ggctgcagct gcaggacgca
      481 ggaccagcac gcacatacaa gggctccctg gattgtctgg cccagatcta tggacacgag
      541 ggactgaggg gagtgaatag gggcatggtg agcaccctgc tgagggagac accctccttc
      601 ggcgtgtact ttctgaccta tgacgcactg acacgggccc tgggatgcga gcccggcgat
      661 agactgctgg tgcctaagct gctgctggca ggaggcacct ccggaatcgt gtcttggctg
      721 agcacatacc ccgtggacgt ggtgaagtct aggctgcagg cagatggact gaggggagca
      781 cctcggtaca gaggcatcct ggattgcgtg caccagtctt atagggcaga gggatggcgg
      841 gtgttcacca gaggactggc aagcacactg ctgagggcat tccctgtgaa cgcagccacc
      901 tttgccaccg tgacagtggt gctgacatat gcccggggcg aggaggccgg cccagagggc
      961 gaggccgtgc ccgccgcccc tgccggccca gccctggccc agcccagctc cctgtaagac
     1021 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     1081 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     1141 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1201 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     1261 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     1321 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1381 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     1441 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1501 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1561 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1621 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1681 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1741 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1801 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     1861 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1921 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1981 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2041 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     2101 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     2161 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2221 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2281 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2341 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2401 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2461 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2521 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2581 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2641 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2701 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2761 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2821 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2881 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2941 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3001 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3061 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3121 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3181 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3241 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3301 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3361 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     3421 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
