LOCUS       sequence-346612-        3783 bp    DNA     circular SYN 15-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 3783)
  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 3783)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SLC35E1_STOP
FEATURES             Location/Qualifiers
     source          1..3783
                     /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(1340..1439)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1456..1475)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1457..1475)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1480..1496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1480..1496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1598..1617)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1609..2418
                     /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(1676..1695)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2565..3153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3054..3073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3307..3324
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3483..3510)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3602..3688)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3737..3759
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3751..3768
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3752..3768
                     /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 agcagcagca
      121 gtgggagcag gacacggcgc cggcggccct ggcgccgcca gctcctctgg cggcgccagg
      181 gagggcgccc gcgtggccgc cctgtgcctg ctgtggtacg cactgagcgc cggaggcaac
      241 gtggtgaaca aagtgatcct gagcgccttc ccatttcccg tgaccgtgtc cctgtgccac
      301 atcctggccc tgtgcgccgg cctgccccct ctgctgaggg catggagggt gccaccagca
      361 cctccagtgt ctggacctgg accaagccca caccctagct ccggaccact gctgccacct
      421 cggttctacc ctagatatgt gctgccactg gccttcggca agtactttgc cagcgtgtcc
      481 gcccacgtgt ctatctggaa ggtgcccgtg agctatgccc acaccgtgaa ggccacaatg
      541 cctatctggg tggtgctgct gagccggatc atcatgaagg agaagcagtc caccaaggtg
      601 tacctgtctc tgatccccat catcagcggc gtgctgctgg caaccgtgac agagctgtcc
      661 tttgacatgt ggggactggt gtctgccctg gcagccacac tgtgcttctc cctgcagaac
      721 atcttttcta agaaggtgct gcgggattcc agaatccacc acctgaggct gctgaatatc
      781 ctgggctgtc acgccgtgtt ctttatgatc ccaacctggg tgctggtgga cctgtccgcc
      841 ttcctggtgt ctagcgatct gacctacgtg tatcagtggc cctggacact gctgctgctg
      901 gccgtgtctg gcttctgtaa ctttgcccag aatgtgatcg cctttagcat cctgaacctg
      961 gtgtcccctc tgtcttatag cgtggccaat gccaccaagc ggatcatggt catcacagtg
     1021 tctctgatca tgctgagaaa cccagtgacc agcacaaatg tgctgggcat gatgaccgcc
     1081 atcctgggcg tgttcctgta caacaagaca aagtatgacg ccaatcagca ggccaggaag
     1141 cacctgctgc ccgtgaccac agccgatctg tcctctaagg agcggcacag atccccactg
     1201 gagaagcccc acaacggcct gctgttcccc cagcacggcg actaccagta tggccgcaac
     1261 aatatcctga cagatcactt tcagtacagc aggcagtcct accctaactc ctattctctg
     1321 aatcgctatg acgtgtaaga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     1381 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1441 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     1501 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     1561 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     1621 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1681 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     1741 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1801 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1861 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     1921 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1981 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     2041 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     2101 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     2161 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2221 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2281 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2341 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2401 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     2461 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     2521 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2581 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2641 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2701 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2761 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2821 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2881 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2941 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3001 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3061 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3121 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3181 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3241 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3301 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3361 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3421 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     3481 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     3541 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     3601 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     3661 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     3721 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     3781 ccc
//
