LOCUS       sequence-357008-        4233 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 4233)
  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 4233)
  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-SLC1A7_STOP
FEATURES             Location/Qualifiers
     source          1..4233
                     /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"
     regulatory      793..802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1790..1889)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1906..1925)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1907..1925)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1930..1946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1930..1946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2048..2067)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2059..2868
                     /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(2126..2145)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3015..3603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3504..3523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3757..3774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3933..3960)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4052..4138)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4187..4209
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4201..4218
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4202..4218
                     /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 gccacacgca
      121 atcctggcaa ggggaagaga cgtgtgcagg agaaacggcc tgctgatcct gtccgtgctg
      181 tctgtgatcg tgggctgtct gctgggcttc tttctgcgga ccaggcgcct gtctccacag
      241 gagatcagct acttccagtt tcccggcgag ctgctgatga gaatgctgaa gatgatgatc
      301 ctgcccctgg tggtgagctc cctgatgagc ggactggcat ccctggatgc caagacctct
      361 agcagactgg gcgtgctgac agtggcttac tatctgtgga ccacattcat ggccgtgatc
      421 gtgggcatct ttatggtgtc tatcatccac ccaggaagcg ccgcacagaa ggagaccaca
      481 gagcagagcg gcaagcccat catgtcctct gccgacgccc tgctggatct gatcaggaac
      541 atgttccctg ccaatctggt ggaggccacc tttaagcagt accgcacaaa gaccacacca
      601 gtggtgaagt cccctaaggt ggcaccagag gaggcaccac ctaggagaat cctgatctac
      661 ggcgtgcagg aggagaacgg ctctcacgtg cagaatttcg cactggacct gaccccacca
      721 cctgaggtgg tgtataagtc cgagcccggc acatctgatg gcatgaacgt gctgggcatc
      781 gtgttcttta gcgccaccat gggcatcatg ctgggaagga tgggcgactc cggagcacct
      841 ctggtgtctt tctgccagtg tctgaatgag tccgtgatga agatcgtggc cgtggccgtg
      901 tggtatttcc cctttggcat cgtgtttctg atcgccggca agatcctgga gatggacgat
      961 cctcgggccg tgggcaagaa gctgggcttc tactccgtga ccgtggtgtg cggactggtg
     1021 ctgcacggac tgtttatcct gcccctgctg tatttcttta tcacaaagaa gaaccctatc
     1081 gtgttcatca ggggaatcct gcaggccctg ctgatcgccc tggccaccag ctcctctagc
     1141 gccaccctgc ctatcacatt caagtgtctg ctggagaaca atcacatcga caggcgcatc
     1201 gcaaggtttg tgctgccagt gggagcaacc atcaatatgg atggaacagc cctgtacgag
     1261 gcagtggcag caatcttcat cgcccaggtg aacaattatg agctggactt tggccagatc
     1321 atcaccatca gcatcaccgc aacagcagca tccatcggag cagcaggcat cccacaggcc
     1381 ggcctggtga ccatggtcat cgtgctgacc agcgtgggac tgccaaccga cgatatcaca
     1441 ctgatcatcg ccgtggactg ggccctggat aggttccgca caatgatcaa cgtgctgggc
     1501 gatgcactgg cagcaggaat catggcacac atctgccgga aggactttgc cagagatacc
     1561 ggcacagaga agctgctgcc ctgtgagacc aagcccgtga gcctgcagga gatcgtggca
     1621 gcccagcaga atggatgcgt gaagagcgtg gcagaggcat ccgagctgac cctgggacca
     1681 acatgtcccc accacgtgcc tgtgcaggtg gagcaggatg aggagctgcc agccgcctct
     1741 ctgaaccact gcaccatcca gatcagcgag ctggagacaa atgtgtaaga cccagctttc
     1801 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     1861 actatcagtc aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg
     1921 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     1981 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     2041 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     2101 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2161 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2221 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2281 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2341 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     2401 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     2461 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     2521 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2581 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     2641 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2701 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2761 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2821 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     2881 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     2941 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     3001 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3061 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3721 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3781 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3841 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     3901 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3961 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     4021 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     4081 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     4141 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     4201 tgtaaaacga cggccagtct taagctcggg ccc
//
