LOCUS       sequence-349395-        4707 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 4707)
  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 4707)
  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-SLC5A3_STOP
FEATURES             Location/Qualifiers
     source          1..4707
                     /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"
     protein_bind    complement(2264..2363)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2380..2399)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2404..2420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2404..2420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2522..2541)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2533..3342
                     /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(2600..2619)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3489..4077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3978..3997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4231..4248
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4407..4434)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4526..4612)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4661..4683
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4675..4692
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4676..4692
                     /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 ccaccatgag ggccgtgctg
      121 gacacagccg atatcgccat cgtggccctg tacttcatcc tggtcatgtg catcggcttc
      181 tttgccatgt ggaagtccaa caggtctacc gtgagcggct attttctggc cggccgcagc
      241 atgacatggg tgaccatcgg cgccagcctg ttcgtgtcca acatcggctc tgagcacttt
      301 atcggactgg caggaagcgg agcagcatcc ggcttcgccg tgggcgcctg ggagtttaat
      361 gcactgctgc tgctgcagct gctgggatgg gtgttcatcc ccatctacat ccgctccggc
      421 gtgtacacaa tgcctgagta tctgtctaag cggttcggcg gccacagaat ccaggtgtac
      481 tttgccgccc tgtccctgat cctgtatatc ttcaccaagc tgagcgtgga cctgtactcc
      541 ggcgccctgt ttatccagga gtctctgggc tggaatctgt atgtgagcgt gatcctgctg
      601 atcggcatga cagccctgct gacagtgacc ggaggactgg tggccgtgat ctatacagac
      661 accctgcagg ccctgctgat gatcatcggc gccctgaccc tgatgatcat cagcatcatg
      721 gagatcggcg gcttcgagga ggtgaagcgg agatacatgc tggcctctcc agatgtgaca
      781 agcatcctgc tgacctataa cctgtctaac accaatagct gtaacgtgag cccaaagaag
      841 gaggccctga agatgctgag gaaccccaca gacgaggatg tgccatggcc aggctttatc
      901 ctgggacaga ccccagcatc cgtgtggtac tggtgcgcag atcaggtcat cgtgcagcgc
      961 gtgctggcag caaagaatat cgcccacgcc aagggcagca cactgatggc cggcttcctg
     1021 aagctgctgc caatgtttat catcgtggtg cccggcatga tctccaggat cctgttcacc
     1081 gacgatatcg cctgcatcaa ccccgagcac tgcatgctgg tgtgcggatc cagggcaggc
     1141 tgttctaata tcgcctaccc tagactggtc atgaagctgg tgccagtggg actgagggga
     1201 ctgatgatgg ccgtgatgat cgccgccctg atgtctgacc tggatagcat cttcaactct
     1261 gccagcacaa tctttaccct ggacgtgtat aagctgatca ggaagtccgc cagctcccgc
     1321 gagctgatga tcgtgggcag aatcttcgtg gcctttatgg tggtcatctc tatcgcctgg
     1381 gtgcctatca tcgtggagat gcagggcggc cagatgtacc tgtatatcca ggaggtggca
     1441 gattacctga ccccacctgt ggccgccctg ttcctgctgg ccatcttttg gaagcggtgt
     1501 aacgagcagg gagccttcta cggaggaatg gcaggatttg tgctgggagc cgtgcggctg
     1561 atcctggcct tcgcctatag agcccccgag tgcgaccagc ctgataatag accaggcttc
     1621 atcaaggaca tccactacat gtatgtggcc acaggcctgt tttgggtgac aggcctgatc
     1681 accgtgatcg tgagcctgct gacaccaccc cctaccaagg agcagatccg gaccacaacc
     1741 ttttggtcta agaagaacct ggtggtgaag gagaattgca gccccaagga ggagccttat
     1801 cagatgcagg agaagagcat cctgagatgt tccgagaaca atgagaccat caaccacatc
     1861 atccccaatg gcaagtccga ggactctatc aagggcctgc agcctgagga tgtgaacctg
     1921 ctggtgacat gcagggagga gggaaatcct gtggcatccc tgggacactc tgaggcagag
     1981 accccagtgg atgcctactc taacggacag gccgccctga tgggagagaa ggagaggaag
     2041 aaggagacag acgatggcgg ccgctattgg aagttcatcg actggttctg cggctttaag
     2101 tccaagtctc tgagcaagag gtccctgcgc gatctgatgg aggaggaggc cgtgtgcctg
     2161 cagatgctgg aggagaccag acaggtgaaa gtgatcctga acatcggcct gttcgccgtg
     2221 tgcagcctgg gcatcttcat gttcgtgtac ttcagcctgt aagacccagc tttcttgtac
     2281 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     2341 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     2401 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2461 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2521 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2581 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2641 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2701 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2761 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2821 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2881 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2941 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     3001 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3061 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3121 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3181 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3241 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3301 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3361 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     3421 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3481 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3541 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3601 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3661 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3721 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3781 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3841 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3901 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3961 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4021 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4081 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4141 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4201 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4261 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4321 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     4381 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     4441 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     4501 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     4561 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     4621 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     4681 acgacggcca gtcttaagct cgggccc
//
