LOCUS       sequence-376747-        4482 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 4482)
  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 4482)
  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-SLC5A5_STOP
FEATURES             Location/Qualifiers
     source          1..4482
                     /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(2039..2138)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2155..2174)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2156..2174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2179..2195)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2179..2195)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2297..2316)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2308..3117
                     /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(2375..2394)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3264..3852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3753..3772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4006..4023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4182..4209)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4301..4387)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4436..4458
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4450..4467
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4451..4467
                     /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 ggccgtggag
      121 acaggagaga ggcccacctt cggcgcatgg gactacggcg tgtttgccct gatgctgctg
      181 gtgtccacag gcatcggact gtgggtggga ctggcaagag gaggccagag gtctgccgag
      241 gatttcttta ccggaggccg gagactggcc gccctgcctg tgggcctgtc cctgtctgcc
      301 agcttcatga gcgccgtgca ggtgctggga gtgccatccg aggcataccg ctatggcctg
      361 aagtttctgt ggatgtgcct gggccagctg ctgaattccg tgctgacagc cctgctgttc
      421 atgccagtgt tttaccggct gggcctgacc tccacatacg agtatctgga gatgaggttc
      481 tctagggccg tgcggctgtg cggcaccctg cagtatatcg tggccacaat gctgtacacc
      541 ggcatcgtga tctatgcccc cgccctgatc ctgaaccagg tgacaggcct ggacatctgg
      601 gccagcctgc tgtccaccgg catcatctgc accttttaca cagcagtggg aggaatgaag
      661 gcagtggtgt ggacagacgt gttccaggtg gtggtcatgc tgtccggatt ttgggtggtg
      721 ctggcaaggg gcgtgatgct ggtgggagga ccccggcagg tgctgaccct ggcccagaac
      781 cacagcagaa tcaatctgat ggacttcaac cctgatccaa ggtcccgcta taccttctgg
      841 acatttgtgg tgggcggcac actggtgtgg ctgagcatgt acggcgtgaa tcaggcacag
      901 gtgcagcggt acgtggcatg tcggaccgag aagcaggcca agctggccct gctgatcaac
      961 caagtgggcc tgtttctgat cgtgagctcc gccgcctgct gtggcatcgt gatgttcgtg
     1021 ttttacaccg actgcgatcc tctgctgctg ggcagaatct ccgcccccga ccagtatatg
     1081 cctctgctgg tgctggacat cttcgaggat ctgccaggag tgcctggact gtttctggca
     1141 tgtgcctact ctggcaccct gagcacagcc tccacctcta tcaatgccat ggccgccgtg
     1201 accgtggagg atctgatcaa gcctcggctg agaagcctgg ccccaaggaa gctggtcatc
     1261 atctccaagg gcctgtctct gatctatgga tccgcctgcc tgacagtggc cgccctgtct
     1321 agcctgctgg gaggaggcgt gctgcagggc tccttcaccg tgatgggcgt gatctctgga
     1381 cctctgctgg gagccttcat cctgggaatg tttctgccag cctgtaacac cccaggcgtg
     1441 ctggcaggac tgggagcagg cctggccctg tctctgtggg tggccctggg agccacactg
     1501 tacccaccta gcgagcagac catgagagtg ctgccatcct ctgccgcaag atgcgtggcc
     1561 ctgagcgtga atgcatccgg actgctggac ccagccctgc tgccagccaa cgatagctcc
     1621 agagccccat ctagcggaat ggacgcaagc aggcccgccc tggccgattc tttctacgcc
     1681 atcagctatc tgtactatgg cgccctgggc accctgacca cagtgctgtg cggcgccctg
     1741 atctcttgtc tgacaggacc aaccaagcgc agcacactgg cacctggact gctgtggtgg
     1801 gacctggcaa ggcagaccgc atctgtggca ccaaaggagg aggtggccat cctggacgat
     1861 aatctggtga agggccctga ggagctgcca acaggcaata agaagccacc cggctttctg
     1921 cccaccaacg aggatagact gttctttctg ggacagaagg agctggaggg agcaggaagc
     1981 tggacacctt gcgtgggaca cgacggagga agggatcagc aggagaccaa cctgtaagac
     2041 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     2101 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     2161 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2221 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     2281 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2341 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2401 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2461 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2521 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2581 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     2641 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2701 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2761 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2821 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2881 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2941 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     3001 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     3061 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     3121 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     3181 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3241 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3301 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3361 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3421 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3481 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3541 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3601 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3661 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3721 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3781 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3841 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3901 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3961 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4021 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4081 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4141 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     4201 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     4261 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     4321 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     4381 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     4441 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
