LOCUS       sequence-333107-        3921 bp    DNA     circular SYN 15-DEC-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 3921)
  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 3921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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-SLC18B1_STOP
FEATURES             Location/Qualifiers
     source          1..3921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1510..1609)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1626..1645)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1627..1645)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1650..1666)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1650..1666)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1768..1787)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1779..2588
                     /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(1846..1865)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2735..3323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3224..3243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3477..3494
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3653..3680)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3772..3858)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3907..3921,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3921,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacac attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccaccatg gaggccctgg gcgacctgga gggaccaagg gcccctggcg
      181 gcgacgatcc tgccggctcc gccggcgaga ccccaggctg gctgtctcgg gagcaggtgt
      241 tcgtgctgat cagcgccgcc tccgtgaacc tgggctccat gatgtgctac tctatcctgg
      301 gcccattctt tcccaaggag gccgagaaga agggcgcctc taacacaatc atcggcatga
      361 tcttcggctg ttttgccctg ttcgagctgc tggccagcct ggtgtttggc aattatctgg
      421 tgcacatcgg cgccaagttc atgtttgtgg caggaatgtt cgtgagcgga ggagtgacca
      481 tcctgttcgg cgtgctggac agggtgcctg atggaccagt gtttatcgcc atgtgcttcc
      541 tggtgcgcgt gatggatgcc gtgtctttcg ccgccgccat gacagccagc tcctctatcc
      601 tggccaaggc ctttcccaac aatgtggcca ccgtgctggg cagcctggag acattctccg
      661 gactgggact gatcctggga ccacctgtgg gcggctttct gtaccagagc ttcggctacg
      721 aggtgccctt tatcgtgctg ggctgcgtgg tgctgctgat ggtgcctctg aacatgtaca
      781 tcctgccaaa ttatgagtct gaccccggcg agcacagctt ttggaagctg atcgccctgc
      841 ctaaagtggg cctgatcgcc ttcgtgatca acagcctgag ctcctgtttc ggctttctgg
      901 atccaaccct gtccctgttt gtgctggaga agttcaatct gcccgccggc tacgtgggcc
      961 tggtgtttct gggcatggcc ctgtcttatg ccatctctag cccactgttc ggcctgctga
     1021 gcgacaagag gccacccctg cgcaagtggc tgctggtgtt cggcaacctg atcacagccg
     1081 gctgctacat gctgctgggc cctgtgccaa tcctgcacat caagagccag ctgtggctgc
     1141 tggtgctgat cctggtggtg tctggcctga gcgccggcat gagcatcatc cccacctttc
     1201 ctgagatcct gtcctgtgcc cacgagaatg gcttcgagga gggactgtcc acactgggac
     1261 tggtgtccgg cctgttttct gccatgtgga gcatcggagc cttcatggga cccaccctgg
     1321 gaggctttct gtatgagaag atcggcttcg agtgggcagc agcaatccag ggactgtggg
     1381 cactgatctc cggactggca atgggactgt tctacctgct ggagtattcc cggagaaagc
     1441 ggtccaagtc tcagaacatc ctgtctaccg aggaggagag aaccacactg ctgcctaatg
     1501 agacataaga cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     1561 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1621 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1681 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1741 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     1801 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1861 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     1921 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1981 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     2041 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     2101 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     2161 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     2221 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     2281 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     2341 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2401 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2461 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2521 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     2581 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     2641 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     2701 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2881 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3481 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     3541 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     3601 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     3661 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     3721 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     3781 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     3841 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
     3901 tgttttccca gtcacgacgt t
//
