LOCUS       sequence-334669-        4149 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 4149)
  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 4149)
  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-SLC1A4_STOP
FEATURES             Location/Qualifiers
     source          1..4149
                     /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(1706..1805)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1822..1841)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1823..1841)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1846..1862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1846..1862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1964..1983)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1975..2784
                     /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(2042..2061)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2931..3519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3420..3439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3673..3690
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3849..3876)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3968..4054)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4103..4125
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4117..4134
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4118..4134
                     /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 gaagagcaac
      121 gagacaaatg gctacctgga ctccgcccag gcaggacctg cagcaggacc aggagcacct
      181 ggcaccgccg ccggcagggc ccggagatgc gccggcttcc tgaggcgcca ggccctggtg
      241 ctgctgaccg tgagcggcgt gctggcagga gcaggactgg gagccgccct gaggggcctg
      301 tccctgtctc gcacacaggt gacctacctg gcctttcccg gggagatgtt gctcagaatg
      361 ctgagaatga tcatcctgcc actggtggtg tgctctctgg tgagcggagc agccagcctg
      421 gacgcctcct gtctgggccg cctgggagga atcgcagtgg catatttcgg cctgaccaca
      481 ctgagcgcct ccgccctggc agtggccctg gcctttatca tcaagccagg aagcggagca
      541 cagacactgc agagctccga cctgggactg gaggattccg gaccacctcc agtgccaaag
      601 gagaccgtgg actccttcct ggatctggcc cggaacctgt ttccttctaa tctggtggtg
      661 gccgccttca gaacatacgc caccgattat aaggtggtga cccagaactc tagctccggc
      721 aatgtgacac acgagaagat ccctatcggc accgagatcg agggcatgaa catcctggga
      781 ctggtgctgt tcgcactggt gctgggagtg gccctgaaga agctgggaag cgagggagag
      841 gacctgatcc ggttctttaa ctctctgaat gaggccacaa tggtgctggt gagctggatc
      901 atgtggtacg tgcctgtggg catcatgttc ctggtgggct ccaagatcgt ggagatgaag
      961 gatatcatcg tgctggtgac ctccctgggc aagtacatct ttgcctctat cctgggccac
     1021 gtgatccacg gcggcatcgt gctgccactg atctatttcg tgtttacacg gaagaatccc
     1081 ttcagatttc tgctgggact gctggcacct ttcgccacag cctttgccac ctgctctagc
     1141 tccgccaccc tgccatctat gatgaagtgt atcgaggaga acaatggcgt ggacaagagg
     1201 atctcccgct tcatcctgcc catcggcgcc acagtgaaca tggatggagc agcaatcttc
     1261 cagtgcgtgg cagccgtgtt tatcgcccag ctgaacaatg tggagctgaa tgccggccag
     1321 atctttacca tcctggtgac agccaccgcc tctagcgtgg gagcagcagg agtgccagca
     1381 ggaggcgtgc tgacaatcgc catcatcctg gaggcaatcg gactgcctac ccacgacctg
     1441 ccactgatcc tggccgtgga ctggatcgtg gataggacca caaccgtggt gaacgtggag
     1501 ggcgatgcac tgggagcagg catcctgcac cacctgaatc agaaggccac caagaagggc
     1561 gagcaggagc tggccgaggt gaaggtggag gccatcccaa actgtaagtc tgaggaggag
     1621 acaagccccc tggtgaccca ccagaatcca gcaggaccag tggcaagcgc cccagagctg
     1681 gagtccaagg agtctgtgct gtaagaccca gctttcttgt acaaagttgg cattataaga
     1741 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1801 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1861 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1921 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1981 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     2041 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2101 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2161 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2221 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2281 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2341 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2401 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2461 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2521 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2581 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2641 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2701 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2761 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2821 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     2881 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2941 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3001 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3061 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3121 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3181 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3241 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3301 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3361 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3421 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3481 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3541 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3601 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3661 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3721 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3781 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     3841 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     3901 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     3961 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     4021 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     4081 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     4141 ctcgggccc
//
