LOCUS       sequence-479320-        4305 bp    DNA     circular SYN 26-JUL-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 4305)
  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 4305)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 26, 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-SLC8B1_STOP
FEATURES             Location/Qualifiers
     source          1..4305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(681..767)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     816..838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     830..847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..962
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2724..2823)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2840..2859)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2841..2859)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2864..2880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2864..2880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2982..3001)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2993..3802
                     /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(3060..3079)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3949..4305,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
      541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
      661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
      781 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
      841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      901 gttgcaacac attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg
      961 ctccaccatg gcaggccgga gactgaacct gagatgggcc ctgtctgtgc tgtgcgtgct
     1021 gctgatggca gagaccgtga gcggaacaag gggaagctcc accggagcac acatctctcc
     1081 ccagtttcct gccagcggcg tgaatcagac acccgtggtg gactgcagaa aggtgtgcgg
     1141 cctgaacgtg tccgacaggt gcgatttcat ccgcaccaat cctgactgtc actctgatgg
     1201 cggctacctg gattatctgg agggcatctt ctgccacttt ccacctagcc tgctgcctct
     1261 ggcagtgacc ctgtacgtgt cctggctgct gtatctgttt ctgatcctgg gcgtgacagc
     1321 cgccaagttc ttttgtccaa acctgtccgc catctctacc acactgaagc tgagccacaa
     1381 cgtggccggc gtgaccttcc tggcctttgg caatggcgcc ccagacatct tctccgccct
     1441 ggtggccttt tctgatccac acacagcagg actggccctg ggcgccctgt tcggagcagg
     1501 cgtgctggtg accacagtgg tggcaggagg aatcaccatc ctgcaccctt tcatggcagc
     1561 atccaggcca ttctttagag acatcgtgtt ttacatggtg gccgtgttcc tgacctttct
     1621 gatgctgttc aggggaaggg tgacactggc atgggcactg ggatacctgg gactgtacgt
     1681 gttttatgtg gtgaccgtga tcctgtgcac atggatctat cagaggcaga ggcgcggctc
     1741 tctgttctgt cctatgccag tgacccccga gatcctgtcc gactctgagg aggatcgggt
     1801 gtctagcaac acaaatagct acgactatgg cgatgagtac agacctctgt tcttttatca
     1861 ggagaccaca gcccagatcc tggtgagggc cctgaatcca ctggactaca tgaagtggcg
     1921 gagaaagagc gcctattgga aggccctgaa ggtgttcaag ctgcccgtgg agtttctgct
     1981 gctgctgacc gtcccagtgg tggaccccga taaggacgat cagaactgga agcgccccct
     2041 gaattgtctg cacctggtca tctcccctct ggtcgtggtg ctgaccctgc agtctggcac
     2101 atacggcgtg tatgagatcg gaggactggt gcccgtgtgg gtggtggtgg tcatcgcagg
     2161 aacagccctg gcatccgtga ccttctttgc cacaagcgat tcccagccac caaggctgca
     2221 ctggctgttc gcctttctgg gctttctgac ctctgccctg tggatcaacg ccgccgccac
     2281 agaggtggtg aatatcctga ggtctctggg cgtggtgttc cgcctgagca acaccgtgct
     2341 gggactgaca ctgctggcat ggggcaatag catcggcgac gccttctccg acttcaccct
     2401 ggcaaggcag ggatacccaa gaatggcctt cagcgcctgc tttggcggca tcatcttcaa
     2461 catcctggtg ggagtgggac tgggatgtct gctgcagatc tctagaagcc acacagaggt
     2521 gaagctggag ccagatggcc tgctggtgtg ggtgctggca ggcgccctgg gactgagcct
     2581 ggtgtttagc ctggtgtccg tgcccctgca gtgcttccag ctgtccaggg tgtacggctt
     2641 ttgtctgctg ctgttctatc tgaattttct ggtggtggcc ctgctgaccg agttcggcgt
     2701 gatccacctg aagtccatgt aagacccagc tttcttgtac aaagttggca ttataagaaa
     2761 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2821 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2881 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2941 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3001 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3061 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3121 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3181 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3241 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3301 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3361 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3421 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3481 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3541 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3601 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3661 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3721 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3781 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3841 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3901 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4081 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaac
//
