LOCUS       sequence-386448-        3867 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 3867)
  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 3867)
  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-CLN3_STOP
FEATURES             Location/Qualifiers
     source          1..3867
                     /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(1424..1523)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1540..1559)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1541..1559)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1564..1580)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1564..1580)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1682..1701)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1693..2502
                     /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(1760..1779)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2649..3237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3138..3157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3391..3408
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3567..3594)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3686..3772)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3821..3843
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3835..3852
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3836..3852
                     /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 ccaccatggg aggatgcgca
      121 ggatcccgga gaaggttctc cgactctgag ggagaggaga ccgtgccaga gcctaggctg
      181 ccactgctgg atcaccaggg agcacactgg aagaacgcag tgggattctg gctgctggga
      241 ctgtgcaaca atttttccta cgtggtcatg ctgtctgccg cccacgacat cctgagccac
      301 aagcgcacct ctggcaatca gagccacgtg gatccaggcc ccacacctat cccacacaac
      361 agctcctctc ggttcgactg caatagcgtg tccaccgcag ccgtgctgct ggcagatatc
      421 ctgcccaccc tggtcatcaa gctgctggca cctctgggac tgcacctgct gccctatagc
      481 cctagagtgc tggtgtccgg aatctgcgca gcaggatcct tcgtgctggt ggccttttct
      541 cacagcgtgg gcaccagcct gtgcggagtg gtgtttgcat ccatcagctc cggactggga
      601 gaggtgacct tcctgtccct gacagccttt taccctaggg ccgtgatctc ttggtggtct
      661 agcggaacag gaggagcagg actgctgggc gccctgtcct atctgggact gacccaggca
      721 ggactgtctc cacagcagac actgctgagc atgctgggaa tccccgccct gctgctggca
      781 tcctacttcc tgctgctgac ctctcctgag gcccaggacc caggaggaga ggaggaggca
      841 gagagcgccg caagacagcc tctgatcagg accgaggccc cagagtctaa gccaggatcc
      901 tctagctccc tgagcctgag ggagcggtgg acagtgttca agggcctgct gtggtacatc
      961 gtgccactgg tggtggtgta cttcgccgag tacttcatca accagggcct gtttgagctg
     1021 ctgttctttt ggaatacctc cctgtctcac gcccagcagt accgctggta tcagatgctg
     1081 tatcaggccg gcgtgttcgc ctctcgctct agcctgcggt gctgtagaat caggtttaca
     1141 tgggccctgg ccctgctgca gtgtctgaac ctggtgttcc tgctggccga cgtgtggttc
     1201 ggctttctgc ccagcatcta cctggtgttt ctgatcatcc tgtatgaggg actgctggga
     1261 ggagcagcat atgtgaacac cttccacaat atcgccctgg agacaagcga cgagcaccgg
     1321 gagtttgcca tggccgccac ctgcatctcc gatacactgg gaatcagcct gtccggactg
     1381 ctggccctgc ctctgcacga cttcctgtgc cagctgagct aagacccagc tttcttgtac
     1441 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1501 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     1561 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1621 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1681 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     1741 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1801 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1861 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     1921 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     1981 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2041 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2101 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2161 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2221 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2281 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2341 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2401 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2461 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     2521 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     2581 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2641 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2701 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2761 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2821 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2881 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2941 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3001 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3061 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3121 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3181 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3241 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3301 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3361 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3421 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3481 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     3541 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     3601 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     3661 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     3721 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     3781 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     3841 acgacggcca gtcttaagct cgggccc
//
