LOCUS       sequence-259013-        5799 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 5799)
  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 5799)
  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_SLC12A7
FEATURES             Location/Qualifiers
     source          1..5799
                     /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(3924..4023)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4040..4059)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4041..4059)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4064..4080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4064..4080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4182..4201)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4193..5002
                     /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(4260..4279)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5149..5737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5638..5657
                     /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 accatgccta ccaacttcac agtggtgcca gtggaggcac acgcagacgg
      721 aggaggcgat gagaccgcag agaggacaga ggcaccagga accccagagg gaccagagcc
      781 cgagcgccct agcccaggcg acggcaaccc acgggagaat tcccccttcc tgaacaatgt
      841 ggaggtggag caggagtcct tctttgaggg caagaatatg gccctgtttg aggaggagat
      901 ggattctaac cctatggtga gctccctgct gaataagctg gccaactata ccaatctgag
      961 ccagggagtg gtggagcacg aggaggacga ggagtcccgg agaagggagg caaaggcacc
     1021 aagaatgggc acattcatcg gcgtgtacct gccctgcctg cagaacatcc tgggcgtgat
     1081 cctgttcctg aggctgacct ggatcgtggg agtggcaggc gtgctggagt cttttctgat
     1141 cgtggccatg tgctgtacct gtacaatgct gacagccatc tctatgagcg ccatcgcaac
     1201 caatggagtg gtgccagcag gaggcagcta ctatatgatc tcccgctctc tgggacctga
     1261 gtttggagga gcagtgggac tgtgcttcta cctgggcacc acatttgccg gcgccatgta
     1321 tatcctgggc accatcgaga tcttcctgac atatatcagc ccaggcgccg ccatctttca
     1381 ggcagaggca gcaggaggag aggcagcagc catgctgcac aacatgcggg tgtacggcac
     1441 ctgcacactg gtgctgatgg ccctggtggt gttcgtgggc gtgaagtatg tgaataagct
     1501 ggccctggtg tttctggcct gcgtggtgct gtctatcctg gccatctacg ccggcgtgat
     1561 caagagcgcc ttcgaccccc ctgatatccc cgtgtgcctg ctgggcaaca gaacactgtc
     1621 tcgccggagc tttgatgctt gcgttaaagc ctatggcatt cacaacaatt ccgccacctc
     1681 tgccctgtgg ggcctgttct gcaatggaag ccagccatcc gccgcatgtg acgagtactt
     1741 catccagaac aatgtgacag agatccaggg aatccctgga gcagcaagcg gcgtgttcct
     1801 ggagaacctg tggtccacct acgcacacgc aggagccttt gtggagaaga agggagtgcc
     1861 ttccgtgcca gtggcagagg agtctagagc aagcgccctg ccttacgtgc tgacagatat
     1921 cgccgcctcc ttcaccctgc tggtgggcat ctattttcca tctgtgaccg gcatcatggc
     1981 cggctccaac aggtctggcg acctgaagga tgcccagaag agcatcccca ccggcacaat
     2041 cctggccatc gtgaccacat ctttcatcta cctgagctgc atcgtgctgt ttggcgcctg
     2101 tatcgagggc gtggtgctga gggacaagtt cggcgaggcc ctgcagggca acctggtcat
     2161 cggaatgctg gcatggccaa gcccttgggt catcgtgatc ggcagcttct tttccacatg
     2221 cggagcagga ctgcagtccc tgaccggagc acctagactg ctgcaggcaa tcgccaggga
     2281 tggcatcgtg ccattcctgc aggtgtttgg acacggcaag gcaaacggag agccaacctg
     2341 ggcactgctg ctgacagtgc tgatctgtga gaccggcatc ctgatcgcct ctctggacag
     2401 cgtggccccc atcctgagca tgttctttct gatgtgctac ctgttcgtga acctggcatg
     2461 tgcagtgcag accctgctga ggacacccaa ttggcgccct cggttcaagt tttatcactg
     2521 gaccctgtcc ttcctgggca tgtctctgtg cctggccctg atgtttatct gttcttggta
     2581 ctatgcactg agcgccatgc tgatcgcagg atgcatctac aagtatatcg agtacagagg
     2641 cgccgagaag gagtggggcg atggcatcag gggcctgtcc ctgaatgcag caagatatgc
     2701 actgctgagg gtggagcacg gaccaccaca cacaaagaac tggagaccac aggtgctggt
     2761 catgctgaat ctggacgcag agcaggcagt gaagcaccca aggctgctgt ccttcacctc
     2821 tcagctgaag gccggcaagg gcctgacaat cgtgggcagc gtgctggagg gcacctatct
     2881 ggataagcac atggaggccc agagagccga ggagaacatc aggagcctga tgtccaccga
     2941 gaagacaaag ggcttttgcc agctggtggt gtctagctcc ctgcgggacg gcatgagcca
     3001 cctgatccag tccgccggac tgggaggcct gaagcacaat acagtgctga tggcctggcc
     3061 cgcctcttgg aagcaggagg ataacccttt cagctggaag aattttgtgg acaccgtgcg
     3121 cgataccaca gcagcacacc aggccctgct ggtggccaag aacgtggact ccttccctca
     3181 gaatcaggag cggtttggcg gcggccacat cgacgtgtgg tggatcgtgc acgatggcgg
     3241 catgctgatg ctgctgccat tcctgctgcg ccagcacaaa gtgtggcgga agtgtagaat
     3301 gaggatcttt accgtggccc aggtggacga taacagcatc cagatgaaga aggatctgca
     3361 gatgttcctg taccacctga gaatctctgc cgaagtggag gtggtggaga tggtggagaa
     3421 tgacatcagc gcctttacct atgagcgcac actgatgatg gagcagcggt cccagatgct
     3481 gaagcagatg cagctgtcta agaacgagca ggagagagag gcccagctga tccacgacag
     3541 gaataccgcc tcccacacag cagcagcagc aaggacccag gcacctccaa cacccgataa
     3601 ggtgcagatg acctggacac gggagaagct gatcgccgag aagtaccgct ctcgggacac
     3661 aagcctgtcc ggcttcaagg acctgttttc catgaagccc gatcagtcta acgtgagaag
     3721 gatgcacacc gccgtgaagc tgaacggcgt ggtgctgaat aagtcccagg acgcccagct
     3781 ggtgctgctg aacatgcctg gaccacctaa gaacaggcag ggcgatgaga attatatgga
     3841 gttcctggag gtgctgacag agggcctgaa tcgggtgctg ctggtgagag gaggaggaag
     3901 agaagtgatc accatctaca gcgacccagc tttcttgtac aaagttggca ttataagaaa
     3961 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     4021 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     4081 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     4141 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     4201 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     4261 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     4321 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     4381 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     4441 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     4501 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     4561 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     4621 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     4681 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     4741 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     4801 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     4861 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     4921 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     4981 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     5041 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     5101 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5281 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5761 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
