LOCUS       sequence-346621-        3603 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 3603)
  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 3603)
  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-SLC35E4_STOP
FEATURES             Location/Qualifiers
     source          1..3603
                     /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"
     regulatory      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1160..1259)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1276..1295)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1277..1295)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1300..1316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1300..1316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1418..1437)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1429..2238
                     /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(1496..1515)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2385..2973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2874..2893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3127..3144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3303..3330)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3422..3508)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3557..3579
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3571..3588
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3572..3588
                     /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 ccaccatgtg caggtgtcca
      121 cctgagcacc acgacggaag gatgaccagc gccgaagtgg gagcagcagc aggaggagca
      181 caggcagccg gcccacccga gtggcctcca ggctcccctc aggccctgag gcagccagga
      241 agggccagag tggcaatggc cgccctggtg tggctgctgg caggagcatc catgagctcc
      301 ctgaacaagt ggatcttcac cgtgcacggc tttggcaggc ctctgctgct gtctgccctg
      361 cacatgctgg tggccgccct ggcatgccac aggggagcac ggagaccaat gccaggagga
      421 acaaggtgtc gcgtgctgct gctgtccctg accttcggca catctatggc atgcggaaat
      481 gtgggactga gggcagtgcc actggatctg gcccagctgg tgaccacaac cacacccctg
      541 tttaccctgg ccctgtctgc cctgctgctg ggcaggcgcc accaccccct gcagctggcc
      601 gccatgggcc ctctgtgcct gggagcagca tgtagcctgg caggagagtt cagaaccccc
      661 cctacaggct gcggctttct gctggcagca acatgtctga ggggcctgaa gagcgtgcag
      721 cagtccgccc tgctgcagga ggagagactg gacgccgtga ccctgctgta cgccacatcc
      781 ctgccttctt tctgcctgct ggcaggagcc gccctggtgc tggaggcagg cgtggcccca
      841 ccccctaccg ccggcgattc ccggctgtgg gcctgcatcc tgctgtcttg tctgctgagc
      901 gtgctgtata acctggcctc tttcagcctg ctggccctga cctctgccct gacagtgcac
      961 gtgctgggca acctgaccgt ggtgggcaat ctgatcctga gccggctgct gtttggcagc
     1021 agactgtccg ccctgtctta cgtgggcatc gccctgacac tgagcggcat gttcctgtat
     1081 cacaattgtg agtttgtggc ctcctgggca gcaaggagag gactgtggag gagggaccag
     1141 ccaagcaagg gcctgtaaga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     1201 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1261 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     1321 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     1381 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     1441 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1501 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     1561 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1621 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1681 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     1741 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1801 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1861 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1921 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     1981 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2041 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2101 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2161 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2221 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     2281 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     2341 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2401 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2461 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2521 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2581 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2641 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2701 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2761 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2821 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2881 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2941 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3001 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3061 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3121 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3181 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3241 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     3301 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     3361 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     3421 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     3481 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     3541 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     3601 ccc
//
