LOCUS       sequence-420914-        4179 bp    DNA     circular SYN 14-APR-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 4179)
  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 4179)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SLC1A3_STOP
FEATURES             Location/Qualifiers
     source          1..4179
                     /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(2304..2403)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2420..2439)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2421..2439)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2444..2460)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2444..2460)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2562..2581)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2573..3382
                     /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(2640..2659)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3529..4117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4018..4037
                     /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 accatgacca agtctaacgg cgaggagcca aagatgggcg gccggatgga
      721 gagattccag cagggcgtgc ggaagagaac cctgctggcc aagaagaagg tgcagaacat
      781 cacaaaggag gacgtgaagt cctacctgtt ccggaatgcc tttgtgctgc tgaccgtgac
      841 agccgtgatc gtgggcacca tcctgggctt tacactgagg ccataccgca tgtcctatcg
      901 ggaggtgaag tatttctctt ttcccggcga gctgctgatg agaatgctgc agatgctggt
      961 gctgcctctg atcatcagct ccctggtgac cggaatggcc gccctggata gcaaggcctc
     1021 cggcaagatg ggcatgagag ccgtggtgta ctatatgacc acaaccatca tcgccgtggt
     1081 catcggcatc atcatcgtga tcatcatcca ccccggcaag ggcaccaagg agaatatgca
     1141 cagggagggc aagatcgtgc gcgtgacagc agcagacgcc ttcctggatc tgatcaggaa
     1201 catgtttccc cctaatctgg tggaggcctg cttcaagcag tttaagacca actacgagaa
     1261 gcggagcttc aaggtgccta tccaggccaa tgagacactg gtgggcgccg tgatcaacaa
     1321 cgtgagcgag gccatggaga ccctgacaag gatcaccgag gagctggtgc cagtgccagg
     1381 atctgtgaac ggcgtgaatg ccctgggcct ggtggtgttc agcatgtgct tcggctttgt
     1441 gatcggcaac atgaaggagc agggccaggc cctgagggag ttctttgaca gcctgaatga
     1501 ggccatcatg cgcctggtgg ccgtgatcat gtggtatgcc cccgtgggca tcctgtttct
     1561 gatcgccggc aagatcgtgg agatggagga tatgggcgtg atcggcggcc agctggccat
     1621 gtacaccgtg acagtgatcg tgggcctgct gatccacgcc gtgatcgtgc tgcctctgct
     1681 gtatttcctg gtgacccgga agaacccatg ggtgtttatc ggaggactgc tgcaggccct
     1741 gatcaccgcc ctgggcacat ctagctcctc tgccaccctg cctatcacat tcaagtgcct
     1801 ggaggagaac aatggcgtgg acaagcgggt gaccagattt gtgctgccag tgggcgccac
     1861 catcaacatg gatggaacag ccctgtacga ggccctggca gcaatcttca tcgcccaggt
     1921 gaacaatttc gagctgaatt ttggccagat catcacaatc tctatcaccg caacagcagc
     1981 aagcatcgga gcagcaggca tcccacaggc cggcctggtg accatggtca tcgtgctgac
     2041 cagcgtggga ctgccaaccg acgatatcac actgatcatc gccgtggact ggtttctgga
     2101 taggctgcgc acaaccacaa acgtgctggg cgacagcctg ggagcaggaa tcgtggagca
     2161 cctgtcccgg cacgagctga agaatagaga tgtggagatg ggcaacagcg tgatcgagga
     2221 gaatgagatg aagaagccct atcagctgat cgcccaggac aacgagaccg agaagcctat
     2281 cgattccgag acaaagatgt aagacccagc tttcttgtac aaagttggca ttataagaaa
     2341 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2401 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2461 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2521 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2581 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2641 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2701 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2761 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2821 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2881 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2941 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3001 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3061 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3121 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3181 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3241 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3301 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3361 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3421 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3481 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3541 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3601 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3661 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3721 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3781 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3841 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3901 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3961 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4021 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4081 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4141 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
