LOCUS       sequence-479327-        4512 bp    DNA     circular SYN 23-JUL-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 4512)
  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 4512)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 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-SLC44A3_STOP
FEATURES             Location/Qualifiers
     source          1..4512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(557..584)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(676..762)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     811..833
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     825..842
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     826..842
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    858..957
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2926..3025)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3042..3061)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3043..3061)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3066..3082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3066..3082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3184..3203)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3195..4004
                     /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(3262..3281)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(4151..4512,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
      541 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
      601 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
      661 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
      721 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      781 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      841 gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
      901 aacacattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctcca
      961 ccatgcactg cctgggcgcc gagtacctgg tgagcgccga gggagcacca aggcagagag
     1021 agtggcggcc tcagatctat agaaagtgta cagacaccgc ctggctgttc ctgttctttc
     1081 tgttttggac cggcctggtg ttcatcatgg gctacagcgt ggtggcagga gcagcaggca
     1141 ggctgctgtt cggctatgac tcctttggca acatgtgcgg caagaagaat tccccagtgg
     1201 agggagcacc actgtctgga caggatatga cactgaagaa gcacgtgttc tttatgaact
     1261 cctgtaatct ggaggtgaag ggcacccagc tgaaccgcat ggccctgtgc gtgtctaatt
     1321 gtccagagga gcagctggac agcctggagg aggtgcagtt ctttgccaac acatctggca
     1381 gcttcctgtg cgtgtacagc ctgaactcct ttaattatac ccactcccca aaggccgatt
     1441 ctctgtgccc taggctgcca gtgccacctt ccaagtcttt tcccctgttc aacagatgcg
     1501 tgccccagac acctgagtgt tactccctgt tcgcctctgt gctgatcaat gacgtggata
     1561 cactgcaccg gatcctgagc ggcatcatgt ccggcagaga caccatcctg ggactgtgca
     1621 tcctggccct ggccctgtct ctggccatga tgttcacctt tcggttcatc accacactgc
     1681 tggtgcacat cttcatcagc ctggtcatcc tgggcctgct gttcgtgtgc ggcgtgctgt
     1741 ggtggctgta ctatgactat acaaacgatc tgtccatcga gctggatacc gagagagaga
     1801 atatgaagtg cgtgctgggc ttcgccatcg tgtctacagg catcaccgcc gtgctgctgg
     1861 tgctgatctt cgtgctgagg aagcgcatca agctgacagt ggagctgttt cagatcacca
     1921 ataaggccat cagctccgcc ccttttctgc tgttccagcc actgtggaca tttgccatcc
     1981 tgatcttctt ttgggtgctg tgggtggccg tgctgctgag cctgggaacc gcaggagcag
     2041 cccaggtcat ggagggcggc caggtggagt acaagcctct gtctggcatc aggtacatgt
     2101 ggagctatca cctgatcggc ctgatctgga catccgagtt catcctggcc tgccagcaga
     2161 tgacaatcgc cggcgccgtg gtgacctgtt actttaaccg ctccaagaat gacccacccg
     2221 atcaccctat cctgtctagc ctgtctatcc tgttctttta tcaccagggc accgtggtga
     2281 agggctcttt cctgatcagc gtggtgcgga tcccaagaat catcgtgatg tacatgcaga
     2341 acgccctgaa ggagcagcag cacggcgccc tgagcaggta cctgttccgc tgctgttatt
     2401 gctgtttttg gtgcctggac aagtacctgc tgcacctgaa ccagaatgcc tataccacaa
     2461 ccgccatcaa tggcacagac ttctgtacct ccgccaagga tgcctttaag atcctgagca
     2521 agaactcctc tcacttcaca tccatcaatt gctttggcga tttcatcatc tttctgggca
     2581 aggtgctggt ggtgtgcttc accgtgtttg gcggcctgat ggccttcaac tacaataggg
     2641 catttcaagt gtgggcagtg ccactgctgc tggtggcctt ctttgcctat ctggtggccc
     2701 actctttcct gagcgtgttt gagacagtgc tggatgccct gttcctgtgc tttgccgtgg
     2761 acctggagac caacgatggc agctccgaga agccttactt catggaccag gagttcctga
     2821 gctttgtgaa gaggtccaac aagctgaaca atgcccgcgc ccagcaggat aagcacagcc
     2881 tgaggaatga ggagggcacc gagctgcagg ccatcgtgcg ctaagaccca gctttcttgt
     2941 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     3001 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     3061 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     3121 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     3181 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     3241 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     3301 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     3361 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     3421 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     3481 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     3541 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     3601 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     3661 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     3721 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     3781 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     3841 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     3901 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     3961 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     4021 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     4081 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4501 agcgaacgac ct
//
