LOCUS       sequence-262708-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4404)
  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 4404)
  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_SLC5A12
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2529..2628)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2645..2664)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2646..2664)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2669..2685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2669..2685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2787..2806)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2798..3607
                     /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(2865..2884)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3754..4342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4243..4262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggagg tgaagaactt cgccgtgtgg gattacgtgg tgtttgccgc
      721 cctgttcttt atcagctccg gcatcggcgt gttctttgcc atcaaggaga ggaagaaggc
      781 cacctccagg gagttcctgg tgggaggccg gcagatgagc tttggccctg tgggactgtc
      841 cctgacagca tccttcatgt ctgccgtgac cgtgctgggc acaccaagcg aggtgtacag
      901 gttcggcgcc tcctttctgg tgttctttat cgcctatctg tttgtgatcc tgctgaccag
      961 cgagctgttc ctgcccgtgt tttaccgcag cggcatcacc tccacatacg agtatctgca
     1021 gctgcggttc aacaagcctg tgagatacgc cgccaccgtg atctatatcg tgcagaccat
     1081 cctgtacaca ggagtggtgg tgtatgcacc tgccctggcc ctgaatcagg tgaccggctt
     1141 cgatctgtgg ggctccgtgt ttgccacagg catcgtgtgc accttttatt gtacactggg
     1201 aggcctgaag gcagtggtgt ggaccgacgc cttccagatg gtggtcatga tcgtgggctt
     1261 tctgaccgtg ctgatccagg gaagcacaca cgcaggagga ttccacaacg tgctggagca
     1321 gtctaccaat ggcagcaggc tgcacatctt cgactttgat gtggacccac tgcggagaca
     1381 cacattctgg accatcacag tgggcggcac ctttacatgg ctgggcatct acggcgtgaa
     1441 ccagtctacc atccagcggt gcatcagctg taagacagag aagcacgcca agctggccct
     1501 gtacttcaat ctgctgggcc tgtggatcat cctggtgtgc gccgtgttct ctggcctgat
     1561 catgtatagc cactttaagg attgtgaccc atggacctcc ggcatcatct ctgcccctga
     1621 ccagctgatg ccatatttcg tgatggagat ctttgccaca atgccaggac tgcctggact
     1681 gttcgtggca tgcgccttta gcggcaccct gtccacagtg gcctctagca tcaatgccct
     1741 ggccaccgtg acattcgagg attttgtgaa gtcctgtttc ccccacctgt ctgacaagct
     1801 gagcacctgg atctccaagg gcctgtgcct gctgtttggc gtgatgtgca catccatggc
     1861 agtggcagca tctgtgatgg gaggagtggt gcaggcaagc ctgtccatcc acggaatgtg
     1921 cggaggacca atgctgggac tgttcagcct gggcatcgtg ttcccctttg tgaactggaa
     1981 gggcgccctg ggaggactgc tgaccggcat cacactgagc ttctgggtgg ccatcggcgc
     2041 ctttatctac ccagcaccag catccaagac ctggcctctg ccactgtcca cagatcagtg
     2101 tatcaagtct aatgtgaccg caacaggacc acctgtgctg tcctctaggc caggaatcgc
     2161 cgacacctgg tactctatca gctatctgta ctattctgcc gtgggatgcc tgggatgtat
     2221 cgtggcaggc gtgatcatca gcctgatcac cggccggcag agaggagagg atatccagcc
     2281 cctgctgatc cggcccgtgt gcaacctgtt ctgtttttgg tccaagaagt acaagaccct
     2341 gtgctggtgt ggcgtgcagc acgactctgg cacagagcag gagaacctgg agaatggctc
     2401 tgccagaaag cagggcgccg agagcgtgct gcagaatgga ctgaggaggg agtccctggt
     2461 gcacgtgcca ggctacgatc ccaaggacaa gtcttataac aatatggcct tcgagaccac
     2521 acactttgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2581 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2641 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2701 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2761 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2821 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2881 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2941 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3001 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3061 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3121 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3181 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3241 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3301 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3361 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3421 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3481 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3541 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3601 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3661 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3721 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3781 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3841 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3901 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3961 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4021 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4081 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4141 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4201 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4261 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4321 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4381 ttgctggcct tttgctcaca tgtt
//
