LOCUS       sequence-334671-        4212 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 4212)
  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 4212)
  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-SLC22A12_STOP
FEATURES             Location/Qualifiers
     source          1..4212
                     /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(1769..1868)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1885..1904)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1886..1904)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1909..1925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1909..1925)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2027..2046)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2038..2847
                     /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(2105..2124)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2994..3582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3483..3502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3736..3753
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3912..3939)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4031..4117)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4166..4188
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4180..4197
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4181..4197
                     /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 ccaccatggc cttttccgag
      121 ctgctggacc tggtgggagg actgggccgg ttccaggtgc tgcagaccat ggccctgatg
      181 gtgagcatca tgtggctgtg cacacagtcc atgctggaga actttagcgc cgcagtgcca
      241 tcccacagat gttgggcccc tctgctggat aattctaccg cccaggccag catcctggga
      301 tctctgagcc ctgaggccct gctggcaatc agcatcccac ctggacctaa ccagaggcca
      361 caccagtgcc ggcggttccg gcagccacag tggcagctgc tggaccccaa tgccaccgcc
      421 acatcctggt ctgaggcaga taccgagcca tgcgtggacg gatgggtgta cgataggagc
      481 atcttcacct ccacaatcgt ggccaagtgg aacctggtgt gcgactctca cgccctgaag
      541 ccaatggccc agagcatcta tctggccggc atcctggtgg gagcagcagc atgcggacca
      601 gcaagcgata ggtttggcag gcgcctggtg ctgacctggt cctacctgca gatggccgtg
      661 atgggaacag cagcagcctt tgccccagca ttccccgtgt actgtctgtt ccgctttctg
      721 ctggccttcg ccgtggcagg cgtgatgatg aacaccggca cactgctgat ggagtggacc
      781 gcagcaaggg caaggccact ggtcatgacc ctgaactctc tgggcttcag ctttggccac
      841 ggactgaccg cagcagtggc atacggcgtg cgggactgga cactgctgca gctggtggtg
      901 tccgtgccct tcttcctgtg cttcctgtat tcctggtggc tggcagagtc tgccaggtgg
      961 ctgctgacca caggccgcct ggactgggga ctgcaggagc tgtggagagt ggcagccatc
     1021 aatggcaagg gcgccgtgca ggataccctg acaccagagg tgctgctgag cgccatgagg
     1081 gaggagctgt ccatgggaca gccaccagca tctctgggca ccctgctgag aatgcctggc
     1141 ctgcggttca gaacctgcat cagcaccctg tgctggttcg cctttggctt cacattcttt
     1201 ggactggccc tggacctgca ggccctggga tctaacatct ttctgctgca gatgttcatc
     1261 ggcgtggtgg atatcccagc aaagatgggc gccctgctgc tgctgtccca cctgggccgg
     1321 agacctaccc tggcagcatc tctgctgctg gcaggactgt gcatcctggc aaatacactg
     1381 gtgccacacg agatgggcgc cctgaggagc gccctggccg tgctgggact gggaggagtg
     1441 ggcgccgcct ttacctgtat cacaatctac agctccgagc tgttccctac cgtgctgagg
     1501 atgacagcag tgggactggg ccagatggca gcaaggggag gagcaatcct gggaccactg
     1561 gtgcgcctgc tgggcgtgca cggcccatgg ctgcccctgc tggtgtatgg caccgtgcct
     1621 gtgctgtctg gactggccgc cctgctgctg ccagagaccc agagcctgcc tctgccagac
     1681 acaatccagg atgtgcagaa ccaggccgtg aagaaggcca cccacggcac actgggcaat
     1741 tccgtgctga agtctaccca gttctaagac ccagctttct tgtacaaagt tggcattata
     1801 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     1861 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     1921 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1981 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     2041 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     2101 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     2161 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2221 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2281 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     2341 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     2401 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     2461 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     2521 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     2581 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     2641 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     2701 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     2761 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     2821 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     2881 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     2941 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3121 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3661 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3721 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3781 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3841 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
     3901 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
     3961 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
     4021 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     4081 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     4141 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt
     4201 aagctcgggc cc
//
