LOCUS       sequence-479343-        3921 bp    DNA     circular SYN 14-AUG-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 3921)
  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 3921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 14, 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-SLC38A6_STOP
FEATURES             Location/Qualifiers
     source          1..3921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(65..151)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     200..222
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     214..231
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     215..231
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    247..346
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1724..1823)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1840..1859)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1841..1859)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1864..1880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1864..1880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1982..2001)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1993..2802
                     /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(2060..2079)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2949..3537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3438..3457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3691..3708
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3867..3894)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
       61 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
      121 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
      181 acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
      241 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acacattgat
      301 gagcaatgct tttttataat gccaactttg tacaaaaaag caggctccac catggaggcc
      361 tcctggggct ctttcaacgc agagagggga tggtacgtga gcgtgcagca gccagaggag
      421 gcagaggcag aggagctgtc tccactgctg agcaatgagc tgcaccggca gagatcccct
      481 ggcgtgtctt tcggcctgag cgtgtttaac ctgatgaatg ccatcatggg ctccggcatc
      541 ctgggcctgg cctatgtgct ggccaacacc ggcgtgtttg gcttctcttt tctgctgctg
      601 acagtggccc tgctggcctc ctactctgtg cacctgctgc tgtctatgtg catccagacc
      661 gccgtgacaa gctatgagga cctgggcctg ttcgcctttg gcctgccagg caagctggtg
      721 gtggcaggaa ccatcatcat ccagaatatc ggcgccatga gctcctacct gctgatcatc
      781 aagaccgagc tgccagcagc aatcgcagag ttcctgacag gcgactattc caggtactgg
      841 tatctggatg gccagaccct gctgatcatc atctgcgtgg gcatcgtgtt cccactggcc
      901 ctgctgccca agatcggctt tctgggctac acatctagcc tgtccttctt tttcatgatg
      961 tttttcgccc tggtggtcat catcaagaag tggtctatcc cctgccctct gaccctgaac
     1021 tatgtggaga agggcttcca gatcagcaat gtgacagacg attgtaagcc taagctgttc
     1081 cactttagca aggagtccgc ctacgccctg ccaaccatgg ccttctcttt tctgtgccac
     1141 acaagcatcc tgcccatcta ttgtgagctg cagagccctt ccaagaagcg catgcagaac
     1201 gtgaccaata cagccatcgc cctgagcttc ctgatctact ttatctccgc cctgttcggc
     1261 tacctgacct tttatgacaa ggtggagagc gagctgctga agggctactc caagtatctg
     1321 tctcacgatg tggtggtcat gaccgtgaag ctgtgcatcc tgttcgccgt gctgctgaca
     1381 gtgcctctga tccactttcc agccaggaag gccgtgacca tgatgttttt cagcaacttc
     1441 cccttttcct ggatccgcca cttcctgatc acactggccc tgaacatcat catcgtgctg
     1501 ctggccatct acgtgcccga tatcaggaac gtgttcggag tggtgggagc atctaccagc
     1561 acatgtctga tcttcatctt tcctggcctg ttctatctga agctgtccag agaggacttt
     1621 ctgtcttgga agaagctggg cgccttcgtg ctgctgatct ttggcatcct ggtgggcaac
     1681 ttctccctgg ccctgatcat ctttgattgg atcaataagt aagacccagc tttcttgtac
     1741 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1801 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     1861 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1921 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1981 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2041 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2101 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2161 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2221 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2281 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2341 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2401 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2461 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2521 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2581 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2641 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2701 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2761 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     2821 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     2881 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2941 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3001 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3061 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3601 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3661 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3721 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3781 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     3841 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     3901 tttgatgcct ggcagtttat g
//
