LOCUS       sequence-433207-        4359 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 4359)
  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 4359)
  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-SLC47A2_STOP
FEATURES             Location/Qualifiers
     source          1..4359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2484..2583)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2600..2619)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2601..2619)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2624..2640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2624..2640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2742..2761)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2753..3562
                     /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(2820..2839)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3709..4297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4198..4217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggact ccctgcagga taccgtggcc ctggaccacg gaggatgctg
      721 tcccgccctg tctaggctgg tgcctagagg attcggcacc gagatgtgga cactgtttgc
      781 cctgtccggc cccctgttcc tgtttcaggt gctgacattc atgatctaca tcgtgtccac
      841 cgtgttttgc ggacacctgg gcaaggtgga gctggcatct gtgaccctgg cagtggcctt
      901 cgtgaacgtg tgcggcgtga gcgtgggagt gggactgagc tccgcctgtg acacactgat
      961 gtctcagagc tttggctccc cgaataaaaa acatgtagga gtcatcctgc agagaggcgc
     1021 cctggtgctg ctgctgtgct gtctgccctg ttgggccctg ttcctgaaca cccagcacat
     1081 cctgctgctg tttaggcagg accctgatgt gagccgcctg acacaggatt acgtgatgat
     1141 cttcatccca ggcctgcccg tgatctttct gtacaacctg ctggccaagt atctgcagaa
     1201 tcagggctgg ctgaagggcc aggaggagga gagccctttc cagaccccag gcctgtccat
     1261 cctgcaccca tcccactctc acctgtctag ggccagcttc cacctgtttc agaagatcac
     1321 ctggccacag gtgctgagcg gagtggtggg caactgcgtg aatggcgtgg ccaactacgc
     1381 cctggtgtcc gtgctgaatc tgggcgtgcg cggctctgcc tatgccaata tcatcagcca
     1441 gttcgcccag accgtgtttc tgctgctgta catcgtgctg aagaagctgc acctggagac
     1501 atgggcagga tggtctagcc agtgcctgca ggactggggc ccattctttt ctctggccgt
     1561 gcccagcatg ctgatgatct gcgtggagtg gtgggcctat gagatcggct ccttcctgat
     1621 gggcctgctg tccgtggtgg atctgtctgc ccaggccgtg atctacgagg tggccaccgt
     1681 gacatatatg atcccactgg gactgagcat cggcgtgtgc gtgcgggtgg gaatggcact
     1741 gggagcagcc gataccgtgc aggcaaagcg gagcgccgtg agcggcgtgc tgagcatcgt
     1801 gggcatctcc ctggtgctgg gcaccctgat cagcatcctg aagaaccagc tgggccacat
     1861 ctttacaaat gacgaggatg tgatcgccct ggtgtcccag gtgctgcccg tgtacagcgt
     1921 gttccacgtg tttgaggcca tctgctgcgt gtacggaggc gtgctgaggg gaacaggcaa
     1981 gcaggccttc ggcgcagcag tgaacgcaat cacctactat atcatcggcc tgccactggg
     2041 catcctgctg acatttgtgg tgaggatgcg catcatggga ctgtggctgg gaatgctggc
     2101 atgcgtgttc ctggcaacag cagcctttgt ggcatacacc gcaaggctgg actggaagct
     2161 ggcagcagag gaggccaaga agcactctgg caggcagcag cagcagaggg cagagagcac
     2221 cgcaacaaga ccaggacctg agaaggccgt gctgtcctct gtggccacag gcagctcccc
     2281 tggcatcacc ctgaccacat atagccggtc cgagtgtcac gtggatttct ttagaacacc
     2341 tgaggaggca cacgcactgt ctgccccaac cagcaggctg tccgtgaagc agctggtcat
     2401 ccggagagga gccgccctgg gagcagcaag cgccaccctg atggtgggcc tgacagtgcg
     2461 gatcctggcc accagacact aagacccagc tttcttgtac aaagttggca ttataagaaa
     2521 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2581 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2641 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2701 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2761 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2821 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2881 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2941 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3001 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3061 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3121 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3181 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3241 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3301 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3361 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3421 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3481 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3541 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3601 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3661 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3721 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3781 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3841 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3901 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3961 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4021 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4081 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4141 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4201 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4261 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4321 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
