LOCUS       sequence-253737-        3894 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 3894)
  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 3894)
  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_SLC52A1
FEATURES             Location/Qualifiers
     source          1..3894
                     /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(2019..2118)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2135..2154)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2136..2154)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2159..2175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2159..2175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2277..2296)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2288..3097
                     /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(2355..2374)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3244..3832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3733..3752
                     /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 accatggcag cacctaccct gggcaggctg gtgctgacac acctgctggt
      721 ggccctgttc ggaatgggaa gctgggcagc agtgaacgga atctgggtgg agctgccagt
      781 ggtggtgaag gacctgcctg agggctggtc cctgccatct tacctgagcg tggtggtggc
      841 cctgggaaat ctgggactgc tggtggtgac cctgtggagg cagctggcac caggcaaggg
      901 agagcaggtg cctatccagg tggtgcaggt gctgtccgtg gtgggcaccg ccctgctggc
      961 cccactgtgg caccacgtgg caccagtggc aggacagctg cactctgtgg ccttcctgac
     1021 actggccctg gtgctggcca tggcctgctg tacctctaac gtgacattcc tgccatttct
     1081 gagccacctg ccccctccat ttctgcggtc tttctttctg ggacagggac tgagcgccct
     1141 gctgccatgc gtgctggccc tggtgcaggg agtgggccgg ctggagtgtc cccctgcccc
     1201 caccaatggc acatctggcc cacccctgga cttcccagag agatttcccg ccagcacctt
     1261 cttttgggcc ctgaccgccc tgctggtgac atccgccgca gccttccggg gactgctgct
     1321 gctgctgcct tccctgccat ctgtgaccac aggaggatcc ggacctgagc tgcagctggg
     1381 atctccagga gcagaggagg aggagaagga ggaggaggag gccctgcctc tgcaggagcc
     1441 tccatctcag gcagcaggaa caatccctgg accagatcca gaggcacacc agctgttcag
     1501 cgcccacgga gcctttctgc tgggactgat ggccttcacc tccgccgtga caaacggcgt
     1561 gctgccctct gtgcagtctt ttagctgcct gccttacggc aggctggcct atcacctggc
     1621 agtggtgctg ggaagcgccg caaatccact ggcatgcttc ctggcaatgg gcgtgctgtg
     1681 ccgcagcctg gcaggactgg tgggcctgtc cctgctgggc atgctgtttg gcgcctacct
     1741 gatggcactg gcaatcctgt ccccttgtcc acctctggtg ggaaccacag caggagtggt
     1801 gctggtggtg ctgagctggg tgctgtgcct gtgcgtgttc agctatgtga aggtggccgc
     1861 cagctccctg ctgcacggag gaggccggcc cgccctgctg gcagccggcg tggccatcca
     1921 agtgggctcc ctgctgggcg ccggcgccat gttcccaccc accagcatct atcacgtgtt
     1981 tcagtccaga aaggactgcg tggatccctg tggccctgac ccagctttct tgtacaaagt
     2041 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2101 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2161 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2221 agataaaata atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2281 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2341 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2401 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2461 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2521 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2581 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2641 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2701 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2761 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2821 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2881 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2941 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3001 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3061 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3121 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3181 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3241 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3301 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3361 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3421 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3481 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3541 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3601 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3661 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3721 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3781 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3841 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
