LOCUS       sequence-355045-        5472 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 5472)
  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 5472)
  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-SLC8A1_STOP
FEATURES             Location/Qualifiers
     source          1..5472
                     /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(3029..3128)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3145..3164)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3146..3164)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3169..3185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3169..3185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3287..3306)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3298..4107
                     /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(3365..3384)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4254..4842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4743..4762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4996..5013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5172..5199)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5291..5377)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5426..5448
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5440..5457
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5441..5457
                     /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 ccaccatgta caacatgcgg
      121 agactgtccc tgtctcctac cttcagcatg ggctttcacc tgctggtgac agtgagcctg
      181 ctgttctccc acgtggatca cgtgatcgcc gagaccgaga tggagggcga gggcaatgag
      241 acaggcgagt gcaccggctc ctactattgt aagaagggcg tgatcctgcc aatctgggag
      301 ccacaggatc cctcttttgg cgacaagatc gccagagcca cagtgtattt cgtggccatg
      361 gtgtacatgt ttctgggcgt gtctatcatc gccgacaggt tcatgagctc catcgaagtg
      421 atcacaagcc aggagaagga gatcaccatc aagaagccaa acggcgagac cacaaagacc
      481 acagtgagaa tctggaacga gacagtgagc aatctgaccc tgatggcact gggatctagc
      541 gccccagaga tcctgctgtc cgtgatcgag gtgtgcggcc acaactttac agccggcgat
      601 ctgggacctt ctaccatcgt gggaagcgcc gccttcaata tgtttatcat catcgccctg
      661 tgcgtgtacg tggtgcctga cggcgagacc agaaagatca agcacctgag ggtgttcttt
      721 gtgaccgccg cctggagcat cttcgcctat acatggctgt acatcatcct gtccgtgatc
      781 tctccaggag tggtggaagt gtgggaggga ctgctgacat tctttttctt tcccatctgc
      841 gtggtgttcg catgggtggc agacaggcgc ctgctgtttt acaagtacgt gtacaagaga
      901 tacagggccg gcaagcagag aggcatgatc atcgagcacg agggcgatag gccctcctct
      961 aagaccgaga tcgagatgga cggcaaggtg gtgaactccc acgtggagaa ttttctggat
     1021 ggcgccctgg tgctggaggt ggatgagagg gaccaggacg atgaggaggc aaggagagag
     1081 atggcccgga tcctgaagga gctgaagcag aagcaccctg acaaggagat cgagcagctg
     1141 atcgagctgg ccaactatca ggtgctgtcc cagcagcaga agtctcgcgc cttctaccgg
     1201 atccaggcaa caaggctgat gaccggagca ggcaatatcc tgaagaggca cgcagcagat
     1261 caggcaagga aggccgtgtc catgcacgag gtgaacacag aggtgaccga gaatgacccc
     1321 gtgtctaaga tcttctttga gcagggcacc tatcagtgcc tggagaactg tggcacagtg
     1381 gccctgacca tcatcaggag gggaggcgat ctgaccaaca cagtgttcgt ggattttcgc
     1441 acagaggacg gaaccgcaaa tgcaggatcc gattacgagt tcacagaggg caccgtggtg
     1501 tttaagcctg gcgacaccca gaaggagatc cgggtgggca tcatcgacga tgacatcttc
     1561 gaggaggatg agaactttct ggtgcacctg tctaatgtga aggtgagctc cgaggcaagc
     1621 gaggacggaa tcctggaggc caaccacgtg tccaccctgg catgcctggg aagcccatcc
     1681 accgccacag tgaccatctt cgatgacgat cacgccggca tcttcacatt tgaggagccc
     1741 gtgacccacg tgtctgagag catcggcatc atggaggtga aggtgctgag aacctctggc
     1801 gccaggggca atgtgatcgt gccatataag acaatcgagg gaaccgcaag gggaggagga
     1861 gaggatttcg aggacacctg tggcgagctg gagtttcaga acgacgagat cgtgaagaca
     1921 atcagcgtga aagtgatcga cgatgaggag tacgagaaga ataagacctt ctttctggag
     1981 atcggagagc caaggctggt ggagatgtcc gagaagaagg ccctgctgct gaatgagctg
     2041 ggcggcttca caatcaccgg caagtatctg ttcggccagc ccgtgtttcg caaggtgcac
     2101 gcccgggagc accctatcct gtctacagtg atcaccatcg ccgatgagta cgacgataag
     2161 cagcctctga ccagcaagga ggaggaggag cggagaatcg ccgagatggg cagaccaatc
     2221 ctgggcgagc acacaaagct ggaagtgatc atcgaggaga gctatgagtt caagtccaca
     2281 gtggacaagc tgatcaagaa gaccaacctg gccctggtgg tgggcacaaa tagctggagg
     2341 gagcagttta tcgaggcaat caccgtgtcc gccggagagg acgatgacga tgacgagtgc
     2401 ggcgaggaga agctgccctc ttgtttcgat tatgtgatgc acttcctgac cgtgttttgg
     2461 aaggtgctgt tcgcctttgt gccccctaca gagtactgga acggctgggc ctgcttcatc
     2521 gtgagcatcc tgatgatcgg cctgctgacc gccttcatcg gcgatctggc ctcccacttt
     2581 ggctgtacca tcggcctgaa ggactctgtg acagccgtgg tgttcgtggc cctgggcaca
     2641 tctgtgcctg acacctttgc cagcaaggtg gccgccaccc aggatcagta tgccgacgcc
     2701 tccatcggca atgtgacagg ctctaacgcc gtgaacgtgt tcctgggaat cggagtggca
     2761 tggagcatcg cagcaatcta ccacgcagca aacggagagc agtttaaggt gagcccaggc
     2821 accctggcct tctccgtgac actgtttacc atcttcgcct ttatcaatgt gggcgtgctg
     2881 ctgtacaggc gccggcctga gatcggagga gagctgggag gaccaaggac agcaaagctg
     2941 ctgacctcct gcctgttcgt gctgctgtgg ctgctgtata tcttcttttc tagcctggag
     3001 gcctactgtc acatcaaggg cttttaagac ccagctttct tgtacaaagt tggcattata
     3061 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     3121 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     3181 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     3241 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     3301 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     3361 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     3421 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     3481 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     3541 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     3601 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     3661 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     3721 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     3781 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     3841 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     3901 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     3961 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     4021 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     4081 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     4141 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     4201 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4381 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4921 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4981 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     5041 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     5101 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
     5161 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
     5221 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
     5281 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     5341 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     5401 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt
     5461 aagctcgggc cc
//
