LOCUS       sequence-352008-        4176 bp    DNA     circular SYN 14-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 4176)
  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 4176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-SLC1A5_STOP
FEATURES             Location/Qualifiers
     source          1..4176
                     /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      104..113
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      898..907
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1733..1832)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1849..1868)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1850..1868)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1873..1889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1873..1889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1991..2010)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2002..2811
                     /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(2069..2088)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2958..3546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3447..3466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3700..3717
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3876..3903)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3995..4081)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4130..4152
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4144..4161
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4145..4161
                     /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 ccaccatggt ggccgacccc
      121 cctagagatt ccaagggact ggcagcagca gagcctacag ccaacggcgg cctggccctg
      181 gccagcatcg aggaccaggg agcagcagca ggaggatact gcggctccag ggatcaggtg
      241 cggagatgtc tgcgcgccaa tctgctggtg ctgctgaccg tggtggccgt ggtggccggc
      301 gtggccctgg gcctgggcgt gtctggagca ggaggcgccc tggccctggg acctgagcgg
      361 ctgagcgcct tcgtgtttcc aggagagctg ctgctgaggc tgctgaggat gatcatcctg
      421 cctctggtgg tgtgcagcct gatcggagga gcagcatccc tggacccagg cgccctgggc
      481 agactgggcg cctgggccct gctgttcttt ctggtgacca cactgctggc cagcgccctg
      541 ggagtgggac tggccctggc cctgcagcca ggcgccgcct ccgccgccat caacgcctct
      601 gtgggcgccg ccggcagcgc cgagaatgcc ccctccaagg aagtcctgga ctcttttctt
      661 gatctggcca ggaacatctt tcctagcaat ctggtgtccg ccgccttccg cagctactcc
      721 accacatatg aggagcggaa catcaccggc acaagagtga aggtgccagt gggacaggag
      781 gtggagggca tgaatatcct gggcctggtg gtgttcgcca tcgtgtttgg agtggccctg
      841 aggaagctgg gaccagaggg agagctgctg atcagattct ttaactcttt caatgaggcc
      901 accatggtgc tggtgagctg gatcatgtgg tacgcccctg tgggcatcat gttcctggtg
      961 gccggcaaga tcgtggagat ggaggacgtg ggactgctgt ttgcaaggct gggcaagtat
     1021 atcctgtgct gtctgctggg acacgcaatc cacggactgc tggtgctgcc cctgatctac
     1081 ttcctgttta caaggaagaa cccctatcgc ttcctgtggg gcatcgtgac ccctctggcc
     1141 acagcctttg gcaccagctc ctctagcgcc accctgcctc tgatgatgaa gtgcgtggag
     1201 gagaacaatg gcgtggccaa gcacatctcc cggttcatcc tgccaatcgg cgccacagtg
     1261 aatatggacg gagccgccct gttccagtgc gtggcagccg tgtttatcgc ccagctgtcc
     1321 cagcagtctc tggattttgt gaagatcatc acaatcctgg tgaccgccac agcctcctct
     1381 gtgggagcag caggaatccc agcaggaggc gtgctgaccc tggcaatcat cctggaggcc
     1441 gtgaacctgc ccgtggatca catctccctg atcctggccg tggactggct ggtggatagg
     1501 tcttgtaccg tgctgaacgt ggagggcgac gcactgggag caggactgct gcagaattac
     1561 gtggatcgga cagagtctag aagcaccgag ccagagctga tccaggtgaa gagcgagctg
     1621 ccactggacc cactgccagt gcctacagag gagggcaatc cactgctgaa gcactatagg
     1681 ggaccagcag gcgatgcaac cgtggcatct gagaaggaga gcgtgatgta agacccagct
     1741 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1801 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1861 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1921 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1981 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     2041 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     2101 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2161 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2221 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     2281 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     2341 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2401 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2461 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2521 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     2581 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2641 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2701 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2761 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2821 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2881 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2941 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3001 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3061 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3121 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3181 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3241 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3301 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3361 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3421 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3601 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3661 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3721 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3781 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3841 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3901 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3961 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     4021 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     4081 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     4141 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
