LOCUS       sequence-312190-        4149 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 4149)
  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 4149)
  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-SLC1A4_STOP
FEATURES             Location/Qualifiers
     source          1..4149
                     /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(2274..2373)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2390..2409)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2391..2409)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2414..2430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2414..2430)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2532..2551)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2543..3352
                     /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(2610..2629)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3499..4087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3988..4007
                     /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 accatggaga agagcaacga gacaaatggc tacctggact ccgcccaggc
      721 aggacctgca gcaggaccag gagcacctgg caccgccgcc ggcagggccc ggagatgcgc
      781 cggcttcctg aggcgccagg ccctggtgct gctgaccgtg agcggcgtgc tggcaggagc
      841 aggactggga gccgccctga ggggcctgtc cctgtctcgc acacaggtga cctacctggc
      901 ctttcccggg gagatgttgc tcagaatgct gagaatgatc atcctgccac tggtggtgtg
      961 ctctctggtg agcggagcag ccagcctgga cgcctcctgt ctgggccgcc tgggaggaat
     1021 cgcagtggca tatttcggcc tgaccacact gagcgcctcc gccctggcag tggccctggc
     1081 ctttatcatc aagccaggaa gcggagcaca gacactgcag agctccgacc tgggactgga
     1141 ggattccgga ccacctccag tgccaaagga gaccgtggac tccttcctgg atctggcccg
     1201 gaacctgttt ccttctaatc tggtggtggc cgccttcaga acatacgcca ccgattataa
     1261 ggtggtgacc cagaactcta gctccggcaa tgtgacacac gagaagatcc ctatcggcac
     1321 cgagatcgag ggcatgaaca tcctgggact ggtgctgttc gcactggtgc tgggagtggc
     1381 cctgaagaag ctgggaagcg agggagagga cctgatccgg ttctttaact ctctgaatga
     1441 ggccacaatg gtgctggtga gctggatcat gtggtacgtg cctgtgggca tcatgttcct
     1501 ggtgggctcc aagatcgtgg agatgaagga tatcatcgtg ctggtgacct ccctgggcaa
     1561 gtacatcttt gcctctatcc tgggccacgt gatccacggc ggcatcgtgc tgccactgat
     1621 ctatttcgtg tttacacgga agaatccctt cagatttctg ctgggactgc tggcaccttt
     1681 cgccacagcc tttgccacct gctctagctc cgccaccctg ccatctatga tgaagtgtat
     1741 cgaggagaac aatggcgtgg acaagaggat ctcccgcttc atcctgccca tcggcgccac
     1801 agtgaacatg gatggagcag caatcttcca gtgcgtggca gccgtgttta tcgcccagct
     1861 gaacaatgtg gagctgaatg ccggccagat ctttaccatc ctggtgacag ccaccgcctc
     1921 tagcgtggga gcagcaggag tgccagcagg aggcgtgctg acaatcgcca tcatcctgga
     1981 ggcaatcgga ctgcctaccc acgacctgcc actgatcctg gccgtggact ggatcgtgga
     2041 taggaccaca accgtggtga acgtggaggg cgatgcactg ggagcaggca tcctgcacca
     2101 cctgaatcag aaggccacca agaagggcga gcaggagctg gccgaggtga aggtggaggc
     2161 catcccaaac tgtaagtctg aggaggagac aagccccctg gtgacccacc agaatccagc
     2221 aggaccagtg gcaagcgccc cagagctgga gtccaaggag tctgtgctgt aagacccagc
     2281 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2341 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2401 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2461 atgttacatt gcacaagata aaataatatc atcatgaaca ataaaactgt ctgcttacat
     2521 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2581 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2641 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2701 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2761 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2821 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2881 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2941 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3001 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3061 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3121 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3181 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3241 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3301 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3361 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3421 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3481 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3541 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3601 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3661 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3721 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3781 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3841 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3901 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3961 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4021 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4081 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4141 tcacatgtt
//
