LOCUS       sequence-333108-        4095 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 4095)
  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 4095)
  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-SLC18A2_STOP
FEATURES             Location/Qualifiers
     source          1..4095
                     /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(1684..1783)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1800..1819)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1801..1819)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1824..1840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1824..1840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1942..1961)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1953..2762
                     /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(2020..2039)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2909..3497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3398..3417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3651..3668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3827..3854)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3946..4032)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4081..4095,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4095,1..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 gcactgagcg aactggcact ggtccggtgg ctgcaggaga
      181 gtcggcggtc taggaagctg attctgttta tcgtctttct ggcactgctg ctggacaaca
      241 tgctgctgac cgtggtggtg cccatcatcc cttcctacct gtattctatc aagcacgaga
      301 agaatgccac cgagatccag acagccaggc ccgtgcacac agccagcatc tccgactctt
      361 tccagtctat ctttagctac tatgataact ccaccatggt gacaggcaat gccacccgcg
      421 acctgacact gcaccagacc gccacacagc acatggtgac caacgccagc gccgtgcctt
      481 ccgattgccc atctgaggac aaggatctgc tgaacgagaa tgtgcaagtg ggcctgctgt
      541 ttgccagcaa ggccaccgtg cagctgatca caaacccttt catcggcctg ctgaccaatc
      601 ggatcggcta ccctatccca atcttcgccg gcttttgcat catgttcgtg agcacaatca
      661 tgttcgcctt tagctcctct tatgccttcc tgctgatcgc acggagcctg cagggaatcg
      721 gaagctcctg ttctagcgtg gcaggaatgg gaatgctggc aagcgtgtac accgacgatg
      781 aggagagagg caatgtgatg ggaatcgccc tgggaggcct ggcaatgggc gtgctggtgg
      841 gccccccttt cggcagcgtg ctgtatgagt ttgtgggcaa gaccgcaccc ttcctggtgc
      901 tggccgccct ggtgctgctg gacggagcca tccagctgtt tgtgctgcag ccctctcggg
      961 tgcagcctga gagccagaag ggcacaccac tgaccacact gctgaaggat ccctacatcc
     1021 tgatcgccgc cggcagcatc tgcttcgcca acatgggaat cgcaatgctg gagccagccc
     1081 tgcccatctg gatgatggag acaatgtgct ccaggaagtg gcagctggga gtggcctttc
     1141 tgcctgccag catctcctac ctgatcggca caaatatctt cggcatcctg gcccacaaga
     1201 tgggcagatg gctgtgcgcc ctgctgggca tgatcatcgt gggcgtgtcc atcctgtgca
     1261 tcccttttgc caagaacatc tatggcctga tcgccccaaa tttcggcgtg ggctttgcca
     1321 tcggcatggt ggactcctct atgatgccaa tcatgggcta cctggtggat ctgaggcacg
     1381 tgagcgtgta cggctccgtg tatgccatcg ccgatgtggc cttctgtatg ggctatgcaa
     1441 tcggaccatc cgccggagga gcaatcgcca aggccatcgg ctttccttgg ctgatgacca
     1501 tcatcggcat catcgacatc ctgttcgccc cactgtgctt ctttctgcgc tctccacccg
     1561 ccaaggagga gaagatggcc atcctgatgg atcacaactg tcccatcaag acaaagatgt
     1621 acactcagaa caacatccag tcctatccaa tcggcgaaga cgaagaaagc gaatcagact
     1681 aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     1741 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
     1801 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     1861 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     1921 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     1981 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2041 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     2101 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2161 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     2221 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     2281 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     2341 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     2401 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     2461 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     2521 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     2581 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     2641 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     2701 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     2761 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     2821 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     2881 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2941 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3001 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3061 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3121 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3181 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3241 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3301 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3361 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3421 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3481 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3541 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3601 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3661 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     3721 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3781 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     3841 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     3901 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     3961 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     4021 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     4081 cccagtcacg acgtt
//
