LOCUS       sequence-337939-        3660 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 3660)
  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 3660)
  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-SLC30A8_STOP
FEATURES             Location/Qualifiers
     source          1..3660
                     /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(1217..1316)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1333..1352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1334..1352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1357..1373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1357..1373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1475..1494)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1486..2295
                     /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(1553..1572)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2442..3030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2931..2950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3184..3201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3360..3387)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3479..3565)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3614..3636
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3628..3645
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3629..3645
                     /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 ccaccatgga gttcctggag
      121 cggacctacc tggtgaacga taaggccgcc aagatgtatg cctttacact ggagtccgtg
      181 gagctgcagc agaagccagt gaataaggac cagtgcccaa gggagaggcc agaggagctg
      241 gagagcggag gcatgtacca ctgtcactcc ggctctaagc ccaccgagaa gggcgccaac
      301 gagtacgcct atgccaagtg gaagctgtgc agcgcctccg ccatctgttt catctttatg
      361 atcgcagagg tggtgggagg acacatcgca ggatctctgg cagtggtgac cgatgcagca
      421 cacctgctga tcgacctgac aagcttcctg ctgagcctgt tttccctgtg gctgagctcc
      481 aagccccctt ccaagcggct gaccttcggc tggcacagag cagagatcct gggcgccctg
      541 ctgtccatcc tgtgcatctg ggtggtgaca ggcgtgctgg tgtatctggc ctgtgagcgg
      601 ctgctgtacc ccgattatca gatccaggcc accgtgatga tcatcgtgtc tagctgcgcc
      661 gtggccgcca atatcgtgct gacagtggtg ctgcaccaga ggtgtctggg ccacaaccac
      721 aaggaggtgc aggcaaatgc atctgtgcgc gcagccttcg tgcacgcact gggcgacctg
      781 tttcagtcta tcagcgtgct gatcagcgcc ctgatcatct acttcaagcc cgagtataag
      841 atcgccgatc ctatctgcac cttcatcttt tctatcctgg tgctggccag caccatcaca
      901 atcctgaagg actttagcat cctgctgatg gagggcgtgc ctaagagcct gaactactcc
      961 ggcgtgaagg agctgatcct ggccgtggat ggcgtgctgt ctgtgcacag cctgcacatc
     1021 tggagcctga ccatgaatca ggtcatcctg tccgcccacg tggcaacagc agcatccaga
     1081 gactctcagg tggtgcggag agagatcgcc aaggccctgt ccaagtcttt caccatgcac
     1141 tccctgacaa tccagatgga gtctcctgtg gaccaggatc cagactgcct gttttgtgag
     1201 gatccatgtg actaagaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     1261 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1321 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     1381 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1441 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1501 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1561 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1621 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1681 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1741 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     1801 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     1861 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1921 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     1981 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2041 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2101 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2161 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2221 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2281 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     2341 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     2401 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2461 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2521 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2581 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2641 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2701 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2761 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2821 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2881 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2941 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3001 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3061 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3121 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3181 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3241 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3301 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     3361 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     3421 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     3481 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     3541 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     3601 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
