LOCUS       sequence-352200-        3507 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 3507)
  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 3507)
  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-SLC25A42_STOP
FEATURES             Location/Qualifiers
     source          1..3507
                     /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"
     regulatory      388..397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1064..1163)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1180..1199)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1181..1199)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1204..1220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1204..1220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1322..1341)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1333..2142
                     /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(1400..1419)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2289..2877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2778..2797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3031..3048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3207..3234)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3326..3412)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3461..3483
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3475..3492
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3476..3492
                     /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 ccaccatggg caacggcgtg
      121 aaggagggac ccgtgcggct gcacgaggac gcagaggccg tgctgagctc ctctgtgagc
      181 tccaagcgcg atcaccggca ggtgctgtct agcctgctga gcggcgccct ggcaggcgcc
      241 ctggcaaaga ccgcagtggc acctctggac agaacaaaga tcatcttcca ggtgtcctct
      301 aagcggttca gcgccaagga ggcctttcgg gtgctgtact atacctacct gaacgagggc
      361 ttcctgtctc tgtggagagg caatagcgcc accatggtga gggtggtgcc ttacgccgcc
      421 atccagtttt ctgcccacga ggagtataag cgcatcctgg gcagctacta tggcttccgg
      481 ggagaggccc tgccaccttg gccaagactg tttgcaggcg ccctggcagg aaccacagca
      541 gcatccctga cctacccact ggatctggtg agagccagga tggccgtgac acctaaggag
      601 atgtatagca acatcttcca cgtgtttatc agaatctcca gggaggaggg cctgaagacc
      661 ctgtaccacg gctttatgcc tacagtgctg ggcgtgatcc catacgccgg cctgagcttc
      721 tttacctatg agacactgaa gtccctgcac agggagtact ctggccggag acagccatat
      781 cccttcgagc gcatgatctt tggagcatgc gcaggactga tcggacagag cgcctcctac
      841 ccactggacg tggtgaggcg ccggatgcag accgcaggag tgacaggata tccaagggca
      901 tctatcgcac gcaccctgag gacaatcgtg agagaggagg gagccgtgag gggactgtat
      961 aagggcctgt ccatgaattg ggtgaagggc cccatcgccg tgggcatctc tttcaccaca
     1021 tttgatctga tgcagatcct gctgcggcac ctgcagagct aagacccagc tttcttgtac
     1081 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1141 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     1201 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1261 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1321 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     1381 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1441 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1501 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     1561 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     1621 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     1681 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     1741 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     1801 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     1861 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     1921 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     1981 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2041 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2101 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     2161 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     2221 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2281 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2341 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2401 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2461 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2521 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2581 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2641 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2701 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2761 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2821 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2881 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2941 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3001 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3061 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3121 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     3181 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     3241 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     3301 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     3361 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     3421 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     3481 acgacggcca gtcttaagct cgggccc
//
