LOCUS       sequence-262669-        4083 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 4083)
  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 4083)
  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_SLC7A7
FEATURES             Location/Qualifiers
     source          1..4083
                     /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(2208..2307)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2324..2343)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2325..2343)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2348..2364)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2348..2364)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2466..2485)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2477..3286
                     /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(2544..2563)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3433..4021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3922..3941
                     /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 accatggtgg actccaccga gtacgaggtg gcctctcagc cagaggtgga
      721 gacaagccca ctgggcgatg gagcatcccc aggacctgag caggtgaagc tgaagaagga
      781 gatcagcctg ctgaacggag tatgcctcat agtgggtaat atgatcggct ctggcatctt
      841 tgtgagcccc aagggcgtgc tgatctactc tgccagcttc ggcctgagcc tggtcatctg
      901 ggcagtgggc ggcctgttct ccgtgtttgg cgccctgtgc tatgcagagc tgggaaccac
      961 aatcaagaag tctggcgcca gctacgccta tatcctggag gccttcggcg gctttctggc
     1021 cttcatccgg ctgtggacat ctctgctgat catcgagcca accagccagg ccatcatcgc
     1081 catcacattc gccaactaca tggtgcagcc cctgtttcct tcctgcttcg ccccctatgc
     1141 agcatctaga ctgctggcag cagcatgcat ctgtctgctg acctttatca attgtgccta
     1201 cgtgaagtgg ggcacactgg tgcaggacat cttcacctat gccaaggtgc tggccctgat
     1261 cgccgtgatc gtggcaggaa tcgtgcggct gggacaggga gcatctacac actttgagaa
     1321 cagcttcgag ggcagctcct ttgccgtggg cgacatcgcc ctggccctgt acagcgccct
     1381 gttctcctat tctggctggg ataccctgaa ttacgtgaca gaggagatca agaacccaga
     1441 gaggaatctg cccctgtcca tcggcatctc tatgcctatc gtgaccatca tctatatcct
     1501 gacaaacgtg gcctactata ccgtgctgga catgagggat atcctggcat ccgacgcagt
     1561 ggcagtgacc tttgccgatc agatctttgg catcttcaac tggatcatcc ctctgagcgt
     1621 ggccctgtcc tgcttcggag gcctgaatgc atccatcgtg gcagcctcta ggctgttctt
     1681 tgtgggaagc agggagggac acctgccaga cgccatctgc atgatccacg tggagaggtt
     1741 tacaccagtg ccctccctgc tgttcaacgg catcatggcc ctgatctacc tgtgcgtgga
     1801 ggatatcttt cagctgatca attactatag cttctcctac tggttctttg tgggcctgtc
     1861 catcgtgggc cagctgtatc tgaggtggaa ggagcctgac aggcctaggc cactgaagct
     1921 gagcgtgttc ttcccaatcg tgttttgcct gtgcaccatc ttcctggtgg ccgtgcccct
     1981 gtacagcgat acaatcaact ccctgatcgg aatcgcaatc gccctgtctg gactgccttt
     2041 ctattttctg atcatccggg tgcctgagca caagagacca ctgtacctgc ggagaatcgt
     2101 gggctctgcc acccggtatc tgcaggtgct gtgcatgagc gtggcagcag agatggacct
     2161 ggaggatgga ggcgagatgc ccaagcagag agatcctaag tccaatgacc cagctttctt
     2221 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2281 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2341 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2401 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2461 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2521 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2581 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2641 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2701 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2761 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2821 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2881 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2941 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3001 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3061 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3121 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3181 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3241 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3301 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3361 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3541 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4081 gtt
//
