LOCUS       sequence-479350-        4326 bp    DNA     circular SYN 03-AUG-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 4326)
  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 4326)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 3, 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-SLC19A1_STOP
FEATURES             Location/Qualifiers
     source          1..4326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     291..310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     544..561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(720..747)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(839..925)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     974..996
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     988..1005
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     989..1005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1021..1120
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2903..3002)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3019..3038)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3020..3038)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3043..3059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3043..3059)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3161..3180)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3172..3981
                     /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(3239..3258)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(4128..4326,1..390)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
       61 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      421 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      481 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
      541 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
      601 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
      661 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
      721 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
      781 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
      841 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      901 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      961 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     1021 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
     1081 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggt gcccagctcc
     1141 cctgcagtgg agaagcaggt gccagtggag ccaggacctg acccagagct gcggtcttgg
     1201 agacacctgg tgtgctacct gtgcttctat ggctttatgg cccagatcag acctggcgag
     1261 agcttcatca ccccatacct gctgggcccc gacaagaact ttacaaggga gcaggtgacc
     1321 aatgagatca cacctgtgct gtcctactct tatctggccg tgctggtgcc agtgttcctg
     1381 ctgaccgatt acctgcgcta tacacccgtg ctgctgctgc agggcctgag cttcgtgagc
     1441 gtgtggctgc tgctgctgct gggacacagc gtggcccaca tgcagctgat ggagctgttc
     1501 tactccgtga ccatggccgc ccggatcgcc tactctagct atatcttttc cctggtgagg
     1561 ccagcaaggt accagagggt ggcaggatat tctagggcag ccgtgctgct gggcgtgttc
     1621 acctcctctg tgctgggaca gctgctggtg accgtgggac gggtgagctt ttccacactg
     1681 aactatatca gcctggcctt cctgacattt tccgtggtgc tggccctgtt cctgaagaga
     1741 cccaagaggt ccctgttctt taacagagac gataggggcc gctgcgagac ctctgccagc
     1801 gagctggaga gaatgaatcc aggacctgga ggcaagctgg gacacgcact gagggtggca
     1861 tgtggcgact ctgtgctggc aaggatgctg agggagctgg gcgatagcct gcggagacca
     1921 cagctgagac tgtggtctct gtggtgggtg tttaacagcg ccggctacta tctggtggtg
     1981 tactatgtgc acatcctgtg gaatgaggtg gaccccacca caaactccgc cagggtgtac
     2041 aatggagcag cagatgcagc aagcaccctg ctgggagcaa tcacatcctt cgccgcaggc
     2101 tttgtgaaga tcagatgggc caggtggagc aagctgctga tcgcaggagt gaccgcaaca
     2161 caggcaggac tggtgttcct gctggcacac acacgccacc ctagctccat ctggctgtgc
     2221 tacgccgcct tcgtgctgtt tcggggcagc tatcagttcc tggtgccaat cgccaccttt
     2281 cagatcgcct ctagcctgtc caaggagctg tgcgccctgg tgttcggcgt gaataccttc
     2341 tttgccacaa tcgtgaagac catcatcacc ttcatcgtga gcgacgtgag aggactggga
     2401 ctgcctgtga ggaagcagtt ccagctgtac tccgtgtatt ttctgatcct gtctatcatc
     2461 tacttcctgg gagcaatgct ggatggactg aggcactgcc agcggggcca ccacccacgc
     2521 cagccacctg cacagggact gcggtccgcc gccgaggaga aggccgccca ggccctgtct
     2581 gtgcaggata agggactggg aggactgcag cctgcacaga gcccaccact gtccccagag
     2641 gactctctgg gagcagtggg accagcatct ctggagcaga ggcagagcga tccctatctg
     2701 gcacaggcac cagcaccaca ggcagcagag tttctgtccc ccgtgaccac accttctcca
     2761 tgcaccctgt gcagcgccca ggcatccgga ccagaggcag cagacgagac atgccctcag
     2821 ctggcagtgc accctccagg cgtgtctaag ctgggcctgc agtgtctgcc tagcgatggc
     2881 gtgcagaacg tgaatcagta agacccagct ttcttgtaca aagttggcat tataagaaag
     2941 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     3001 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     3061 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     3121 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     3181 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     3241 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     3301 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     3361 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     3421 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     3481 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     3541 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     3601 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     3661 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     3721 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     3781 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     3841 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     3901 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     3961 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     4021 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     4081 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4141 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4201 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4261 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4321 gcaccg
//
