LOCUS       sequence-362755-        4257 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 4257)
  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 4257)
  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-SLC11A2_STOP
FEATURES             Location/Qualifiers
     source          1..4257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1814..1913)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1930..1949)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1931..1949)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1954..1970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1954..1970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2072..2091)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2083..2892
                     /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(2150..2169)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3039..3627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3528..3547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3781..3798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3957..3984)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4076..4162)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4211..4233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4225..4242
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4226..4242
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggt gctgggcccc
      121 gagcagaaga tgagcgacga ttccgtgtct ggcgaccacg gagagagcgc ctccctggga
      181 aacatcaatc cagcctactc taacccctct ctgagccagt cccccggcga tagcgaggag
      241 tactttgcca catatttcaa tgagaagatc agcatccctg aggaggagta cagctgcttc
      301 tcctttcgga agctgtgggc ctttaccggc cctggcttcc tgatgtccat cgcctatctg
      361 gacccaggca acatcgagtc cgatctgcag tctggagcag tggcaggctt caagctgctg
      421 tggatcctgc tgctggccac actggtgggc ctgctgctgc agaggctggc agcaaggctg
      481 ggagtggtga ccggactgca cctggcagag gtgtgccaca ggcagtaccc caaggtgcct
      541 cgcgtgatcc tgtggctgat ggtggagctg gccatcatcg gcagcgacat gcaggaagtg
      601 atcggctccg ccatcgccat caatctgctg tctgtgggaa ggatccctct gtggggaggc
      661 gtgctgatca caatcgccga caccttcgtg tttctgttcc tggataagta tggcctgcgc
      721 aagctggagg ccttctttgg ctttctgatc acaatcatgg ccctgacctt cggctacgag
      781 tatgtgacag tgaagccatc tcagagccag gtgctgaagg gcatgttcgt gccatcctgc
      841 tctggatgta ggaccccaca gatcgagcag gcagtgggaa tcgtgggagc cgtgatcatg
      901 ccccacaaca tgtacctgca cagcgccctg gtgaagtccc ggcaggtgaa cagaaacaat
      961 aagcaggagg tgagagaggc caataagtat ttctttatcg agtcctgcat cgccctgttc
     1021 gtgagcttca tcatcaacgt gttcgtggtg agcgtgttcg ccgaggcctt ctttggcaag
     1081 acaaacgagc aggtggtgga ggtgtgcacc aatacaagct ccccccacgc cggcctgttt
     1141 cctaaggaca attccaccct ggccgtggat atctacaagg gcggcgtggt gctgggctgt
     1201 tacttcggcc cagccgccct gtatatctgg gcagtgggaa tcctggcagc aggacagtct
     1261 agcaccatga caggcaccta tagcggccag tttgtgatgg agggcttcct gaatctgaag
     1321 tggagccggt ttgccagagt ggtgctgaca aggtccatcg ccatcatccc taccctgctg
     1381 gtggccgtgt ttcaggacgt ggagcacctg accggcatga acgatttcct gaatgtgctg
     1441 cagagcctgc agctgccttt tgccctgatc ccaatcctga cattcacctc tctgcggcca
     1501 gtgatgagcg actttgcaaa cggactggga tggagaatcg caggaggcat cctggtgctg
     1561 atcatctgct ccatcaatat gtacttcgtg gtggtgtatg tgagggatct gggacacgtg
     1621 gccctgtacg tggtggcagc agtggtgtcc gtggcatacc tgggcttcgt gttctacctg
     1681 ggctggcagt gtctgatcgc cctgggcatg tctttcctgg attgcggcca cacatgtcac
     1741 ctgggactga ccgcacagcc tgagctgtat ctgctgaaca ccatggacgc cgattctctg
     1801 gtgagccgct aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1861 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1921 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1981 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2041 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     2101 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2161 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2221 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2281 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2341 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2401 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2461 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2521 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2581 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     2641 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2701 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2761 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2821 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2881 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2941 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     3001 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3061 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3181 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3721 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3781 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3841 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3901 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3961 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     4021 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     4081 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     4141 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     4201 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
