LOCUS       sequence-390326-        3447 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 3447)
  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 3447)
  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-SLC25A6_STOP
FEATURES             Location/Qualifiers
     source          1..3447
                     /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(1004..1103)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1120..1139)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1121..1139)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1144..1160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1144..1160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1262..1281)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1273..2082
                     /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(1340..1359)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2229..2817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2718..2737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2971..2988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3147..3174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3266..3352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3401..3423
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3415..3432
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3416..3432
                     /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 ccaccatgac cgagcaggcc
      121 atcagcttcg ccaaggattt tctggcagga ggaatcgcag cagcaatctc caagacagca
      181 gtggccccca tcgagagagt gaagctgctg ctgcaggtgc agcacgcctc caagcagatc
      241 gccgccgaca agcagtataa gggcatcgtg gattgcatcg tgaggatccc aaaggagcag
      301 ggcgtgctgt ctttctggag aggcaacctg gccaatgtga tcaggtattt tcccacccag
      361 gccctgaact tcgcctttaa ggataagtac aagcagatct tcctgggcgg cgtggacaag
      421 cacacccagt tctggaggta ttttgccggc aatctggcat ctggaggagc agcaggagcc
      481 acaagcctgt gcttcgtgta ccctctggat tttgcaagga ccaggctggc agcagacgtg
      541 ggcaagtctg gcacagagag agagtttagg ggcctgggcg attgtctggt gaagatcacc
      601 aagagcgacg gcatccgcgg cctgtatcag ggcttcagcg tgtccgtgca gggcatcatc
      661 atctaccggg ccgcctattt tggcgtgtac gacaccgcca agggcatgct gcccgatcct
      721 aagaacacac acatcgtggt gagctggatg atcgcacaga ccgtgacagc agtggcagga
      781 gtggtgtcct atccattcga tacagtgcgg agaaggatga tgatgcagtc tggccgcaag
      841 ggcgccgaca tcatgtacac cggcacagtg gattgttggc gcaagatctt ccgggacgag
      901 ggcggcaagg ccttctttaa gggcgcctgg tccaatgtgc tgaggggaat gggaggagcc
      961 tttgtgctgg tgctgtacga cgagctgaag aaagtgatct aagacccagc tttcttgtac
     1021 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1081 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     1141 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1201 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1261 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     1321 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1381 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1441 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     1501 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     1561 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     1621 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     1681 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     1741 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     1801 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     1861 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     1921 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     1981 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2041 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     2101 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     2161 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2221 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2281 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2341 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2401 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2461 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2521 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2581 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2641 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2701 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2761 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2821 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2881 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2941 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3001 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3061 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     3121 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     3181 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     3241 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     3301 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     3361 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     3421 acgacggcca gtcttaagct cgggccc
//
