LOCUS       sequence-405768-        4407 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 4407)
  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 4407)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4407
                     /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(1964..2063)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2080..2099)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2081..2099)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2104..2120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2104..2120)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2222..2241)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2233..3042
                     /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(2300..2319)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3189..3777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3678..3697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3931..3948
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4107..4134)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4226..4312)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4361..4383
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4375..4392
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4376..4392
                     /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 ggtgaagaac
      121 ttcgccgtgt gggattacgt ggtgtttgcc gccctgttct ttatcagctc cggcatcggc
      181 gtgttctttg ccatcaagga gaggaagaag gccacctcca gggagttcct ggtgggaggc
      241 cggcagatga gctttggccc tgtgggactg tccctgacag catccttcat gtctgccgtg
      301 accgtgctgg gcacaccaag cgaggtgtac aggttcggcg cctcctttct ggtgttcttt
      361 atcgcctatc tgtttgtgat cctgctgacc agcgagctgt tcctgcccgt gttttaccgc
      421 agcggcatca cctccacata cgagtatctg cagctgcggt tcaacaagcc tgtgagatac
      481 gccgccaccg tgatctatat cgtgcagacc atcctgtaca caggagtggt ggtgtatgca
      541 cctgccctgg ccctgaatca ggtgaccggc ttcgatctgt ggggctccgt gtttgccaca
      601 ggcatcgtgt gcacctttta ttgtacactg ggaggcctga aggcagtggt gtggaccgac
      661 gccttccaga tggtggtcat gatcgtgggc tttctgaccg tgctgatcca gggaagcaca
      721 cacgcaggag gattccacaa cgtgctggag cagtctacca atggcagcag gctgcacatc
      781 ttcgactttg atgtggaccc actgcggaga cacacattct ggaccatcac agtgggcggc
      841 acctttacat ggctgggcat ctacggcgtg aaccagtcta ccatccagcg gtgcatcagc
      901 tgtaagacag agaagcacgc caagctggcc ctgtacttca atctgctggg cctgtggatc
      961 atcctggtgt gcgccgtgtt ctctggcctg atcatgtata gccactttaa ggattgtgac
     1021 ccatggacct ccggcatcat ctctgcccct gaccagctga tgccatattt cgtgatggag
     1081 atctttgcca caatgccagg actgcctgga ctgttcgtgg catgcgcctt tagcggcacc
     1141 ctgtccacag tggcctctag catcaatgcc ctggccaccg tgacattcga ggattttgtg
     1201 aagtcctgtt tcccccacct gtctgacaag ctgagcacct ggatctccaa gggcctgtgc
     1261 ctgctgtttg gcgtgatgtg cacatccatg gcagtggcag catctgtgat gggaggagtg
     1321 gtgcaggcaa gcctgtccat ccacggaatg tgcggaggac caatgctggg actgttcagc
     1381 ctgggcatcg tgttcccctt tgtgaactgg aagggcgccc tgggaggact gctgaccggc
     1441 atcacactga gcttctgggt ggccatcggc gcctttatct acccagcacc agcatccaag
     1501 acctggcctc tgccactgtc cacagatcag tgtatcaagt ctaatgtgac cgcaacagga
     1561 ccacctgtgc tgtcctctag gccaggaatc gccgacacct ggtactctat cagctatctg
     1621 tactattctg ccgtgggatg cctgggatgt atcgtggcag gcgtgatcat cagcctgatc
     1681 accggccggc agagaggaga ggatatccag cccctgctga tccggcccgt gtgcaacctg
     1741 ttctgttttt ggtccaagaa gtacaagacc ctgtgctggt gtggcgtgca gcacgactct
     1801 ggcacagagc aggagaacct ggagaatggc tctgccagaa agcagggcgc cgagagcgtg
     1861 ctgcagaatg gactgaggag ggagtccctg gtgcacgtgc caggctacga tcccaaggac
     1921 aagtcttata acaatatggc cttcgagacc acacactttt aagacccagc tttcttgtac
     1981 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     2041 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     2101 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2161 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2221 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2281 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2341 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2401 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2461 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2521 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2581 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2641 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2701 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2761 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2821 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2881 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2941 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3001 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3061 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     3121 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3181 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3241 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3301 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3361 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3421 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3481 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3541 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3601 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3661 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3721 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3781 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3841 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3901 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3961 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4021 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     4081 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     4141 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     4201 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     4261 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     4321 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     4381 acgacggcca gtcttaagct cgggccc
//
