LOCUS       sequence-262938-        4233 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 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_SLC38A9
FEATURES             Location/Qualifiers
     source          1..4233
                     /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(2358..2457)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2474..2493)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2475..2493)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2498..2514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2498..2514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2616..2635)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2627..3436
                     /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(2694..2713)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3583..4171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4072..4091
                     /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 accatggcca acatgaatag cgactccagg cacctgggca ccagcgaggt
      721 ggaccacgag agggatccag gacccatgaa catccagttt gagcctagcg atctgcggtc
      781 caagagacca ttctgcatcg agcccaccaa catcgtgaac gtgaatcacg tgatccagcg
      841 ggtgtctgac cacgccagcg ccatgaataa gcggatccac tactattcca gactgaccac
      901 accagcagac aaggccctga tcgcaccaga tcacgtggtg cctgcaccag aggagtgtta
      961 cgtgtattcc cccctgggct ctgcctacaa gctgcagtct tacaccgagg gctatggcaa
     1021 gaacaccagc ctggtgacaa tctttatgat ctggaatacc atgatgggca catccatcct
     1081 gtctatccct tggggcatca agcaggccgg cttcaccaca ggcatgtgcg tgatcatcct
     1141 gatgggcctg ctgaccctgt actgctgtta tagggtggtg aagagccgca caatgatgtt
     1201 ttctctggac accacaagct gggagtaccc cgacgtgtgc aggcactatt ttggctcctt
     1261 cggccagtgg agctccctgc tgttcagcct ggtgtccctg atcggcgcca tgatcgtgta
     1321 ctgggtgctg atgagcaact tcctgtttaa taccggcaag ttcatcttta acttcatcca
     1381 ccacatcaat gacaccgata caatcctgag cacaaacaat tccaaccccg tgatctgccc
     1441 ttctgccgga agcggaggac accctgacaa ctctagcatg atcttttacg ccaatgatac
     1501 cggcgcccag cagtttgaga agtggtggga taagagccgc acagtgccct tctacctggt
     1561 gggcctgctg ctgcccctgc tgaactttaa gtctcctagc ttctttagca agttcaatat
     1621 cctgggcacc gtgtccgtgc tgtacctgat ctttctggtg acattcaagg ccgtgaggct
     1681 gggcttccac ctggagtttc actggttcat ccctaccgag ttctttgtgc cagagatccg
     1741 ctttcagttc ccacagctga ccggcgtgct gacactggcc ttctttatcc acaattgcat
     1801 catcacactg ctgaagaaca ataagaagca ggagaacaat gtgagggacc tgtgcatcgc
     1861 ctatatgctg gtgaccctga catacctgta tatcggcgtg ctggtgtttg cctccttccc
     1921 ctctccccct ctgtccaagg actgtatcga gcagaacttt ctggacaatt tcccttcctc
     1981 tgataccctg tctttcatcg cccggatctt tctgctgttc cagatgatga cagtgtaccc
     2041 actgctgggc tatctggcca gagtgcagct gctgggccac atctttggcg acatctaccc
     2101 ctctatcttc cacgtgctga tcctgaacct gatcatcgtg ggcgccggcg tgatcatggc
     2161 ctgcttctac cccaatatcg gaggaatcat caggtattcc ggagcagcat gtggactggc
     2221 cttcgtgttc atctaccctt ccctgatcta tatcatctct ctgcaccagg aggagagact
     2281 gacctggcca aagctgatct ttcacgtgtt catcatcatc ctgggcgtgg ccaacctgat
     2341 cgtgcagttc tttatggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2401 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2461 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2521 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2581 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2641 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2701 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2761 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2821 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2881 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2941 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3001 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3061 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3121 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3181 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3241 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3301 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3361 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3421 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3481 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3541 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3721 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3901 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3961 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4021 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4081 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4141 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4201 cctggccttt tgctggcctt ttgctcacat gtt
//
