LOCUS       sequence-262893-        4089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4089)
  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 4089)
  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_SLC41A1
FEATURES             Location/Qualifiers
     source          1..4089
                     /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(2214..2313)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2330..2349)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2331..2349)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2354..2370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2354..2370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2472..2491)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2483..3292
                     /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(2550..2569)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3439..4027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3928..3947
                     /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 accatgagct ccaagccaga gcccaaggat gtgcaccagc tgaacggaac
      721 cggaccatct gccagccctt gctctagcga cggaccagga agggagccac tggcaggcac
      781 atctgagttc ctgggaccag atggagcagg agtggaggtg gtcatcgaga gcagggcaaa
      841 tgcaaagggc gtgcgcgagg aggacgcact gctggagaac ggctcccagt ctaatgagtc
      901 cgacgacgtg agcaccgata ggggacctgc accaccttcc ccactgaagg agacaagctt
      961 ctccatcggc ctgcaggtgc tgttcccttt tctgctggca ggatttggaa ccgtggcagc
     1021 aggaatggtg ctggatatcg tgcagcactg ggaggtgttc cagaaggtga cagaggtgtt
     1081 tatcctggtg ccagccctgc tgggcctgaa gggcaacctg gagatgaccc tggcatccag
     1141 gctgtctaca gcagccaata tcggccacat ggataccccc aaggagctgt ggagaatgat
     1201 cacaggcaac atggccctga tccaggtgca ggcaaccgtg gtgggcttcc tggcaagcat
     1261 cgcagccgtg gtgtttggct ggatccctga cggccacttc tccatcccac acgcctttct
     1321 gctgtgcgcc tcctctgtgg ccacagcctt catcgcctcc ctggtgctgg gcatgatcat
     1381 gatcggcgtg atcatcggct ctcggaagat cggcatcaac cccgataatg tggcaacccc
     1441 tatcgcagca agcctgggcg acctgatcac actggccctg ctgagcggaa tctcctgggg
     1501 actgtacctg gagctgaatc actggaggta catctatcct ctggtgtgcg ccttctttgt
     1561 ggccctgctg ccagtgtggg tggtgctggc acggagatct ccagcaacca gggaggtgct
     1621 gtatagcgga tgggagccag tgatcatcgc aatggcaatc agctccgtgg gaggactgat
     1681 cctggataag accgtgagcg accctaactt cgccggcatg gccgtgttta caccagtgat
     1741 caacggagtg ggaggaaatc tggtggcagt gcaggcatct aggatcagca cctttctgca
     1801 catgaacggc atgcccggcg agaattccga gcaggcacct aggcgctgcc ctagcccatg
     1861 taccacattc ttttcccccg acgtgaattc tcggagcgcc agagtgctgt tcctgctggt
     1921 ggtgcctggc cacctggtgt ttctgtacac aatcagctgt atgcagggcg gccacaccac
     1981 actgaccctg atcttcatca tcttttacat gacagccgcc ctgctgcagg tgctgatcct
     2041 gctgtatatc gccgactgga tggtgcactg gatgtgggga aggggactgg acccagataa
     2101 cttctccatc ccctatctga ccgccctggg cgatctgctg ggaacaggac tgctggccct
     2161 gtcttttcac gtgctgtggc tgatcggcga cagagatacc gacgtgggcg acgacccagc
     2221 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2281 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2341 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2401 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2461 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2521 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2581 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2641 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2701 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2761 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2821 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2881 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2941 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3001 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3061 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3121 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3181 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3241 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3301 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3361 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3421 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3481 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3541 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3601 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3661 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3721 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3781 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3841 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3901 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3961 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4021 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4081 tcacatgtt
//
