LOCUS       sequence-351776-        4530 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 4530)
  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 4530)
  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-SLC5A4_STOP
FEATURES             Location/Qualifiers
     source          1..4530
                     /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(2087..2186)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2203..2222)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2204..2222)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2227..2243)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2227..2243)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2345..2364)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2356..3165
                     /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(2423..2442)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3312..3900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3801..3820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4054..4071
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4230..4257)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4349..4435)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4484..4506
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4498..4515
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4499..4515
                     /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 ctccaccgtg
      121 agcccatcca caatcgcaga gaccccagag ccacctccac tgtccgacca catcagaaac
      181 gccgccgata tctctgtgat cgtgatctac ttcctggtgg tcatggcagt gggactgtgg
      241 gccatgctga agacaaatag aggcaccatc ggcggcttct ttctggcagg aagggatatg
      301 gcatggtggc caatgggcgc ctctctgttt gccagcaaca tcggctccaa tcactatgtg
      361 ggactggcag gaacaggagc agcaagcggc gtggccacag tgaccttcga gtggaccagc
      421 tccgtgatgc tgctgatcct gggctggatc tttgtgccaa tctacatcaa gtccggcgtg
      481 atgaccatgc ccgagtacct taaaaagcgc ttcggaggag agcgcctgca ggtgtacctg
      541 agcatcctgt ccctgtttat ctgcgtggtg ctgctgatca gcgccgacat cttcgccggc
      601 gccatcttta tcaagctggc cctgggcctg gatctgtatc tggccatctt catcctgctg
      661 gccatgacag ccgtgtacac cacaaccgga ggactggcat ctgtgatcta tacagacacc
      721 ctgcagacca tcatcatgct gatcggcagc ttcatcctga tgggcttcgc ctttaacgaa
      781 gtgggcggct acgagtcttt tacagagaag tatgtgaacg ccacccctag cgtggtggag
      841 ggcgacaatc tgacaatctc tgccagctgc tacaccccac gcgccgattc cttccacatc
      901 tttcgggacg cagtgaccgg cgatatccca tggccaggca tcatcttcgg catgcccatc
      961 acagccctgt ggtattggtg taccaatcag gtcatcgtgc agcggtgcct gtgcggcaag
     1021 gatatgagcc acgtgaaggc cgcctgcatc atgtgcgcct acctgaagct gctgcctatg
     1081 tttctgatgg tcatgccagg catgatcagc cggatcctgt ataccgacat ggtggcctgc
     1141 gtggtgcctt ccgagtgcgt gaagcactgt ggcgtggatg tgggctgtac aaactacgcc
     1201 tatcccacca tggtgctgga gctgatgcct cagggcctga gaggcctgat gctgtccgtg
     1261 atgctggcct ctctgatgtc tagcctgaca tccatcttca attccgcctc tacactgttt
     1321 accatcgacc tgtacaccaa gatgcgcaag caggcctctg agaaggagct gctgatcgcc
     1381 ggccggatct tcgtgctgct gctgacagtg gtgagcatcg tgtgggtgcc cctggtgcag
     1441 gtgtcccaga acggccagct gatccactac accgagtcca tctcctctta tctgggacca
     1501 cctatcgcag ccgtgttcgt gctggccatc ttttgcaaga gagtgaatga gcagggagca
     1561 ttctggggac tgatggtggg actggcaatg ggactgatca ggatgatcac agagtttgca
     1621 tacggaaccg gaagctgtct ggcaccatcc aactgcccta agatcatctg tggcgtgcac
     1681 tacctgtatt tttctatcgt gctgttcttt ggcagcatgc tggtgacact gggcatcagc
     1741 ctgctgacca agcctatccc agacgtgcac ctgtaccggc tgtgctgggt gctgagaaat
     1801 tccacagagg agaggatcga catcgatgcc gaggagaagt ctcaggagga gaccgacgat
     1861 ggcgtggagg aggactaccc cgagaagtct aggggctgcc tgaagaaggc ctatgatctg
     1921 ttctgtggcc tgcagaaggg ccctaagctg acaaaggagg aggaggaggc cctgagcaag
     1981 aagctgacag atacctctga gcggccaagc tggagaacca tcgtgaacat caatgccatc
     2041 ctgctgctgg ccgtggtggt gtttatccac ggctactatg cctaagaccc agctttcttg
     2101 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     2161 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     2221 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2281 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2341 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2401 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2461 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2521 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2581 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2641 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2701 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     2761 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     2821 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2881 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2941 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     3001 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3061 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3121 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     3181 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     3241 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3301 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3361 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3421 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3481 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3541 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3601 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3661 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3721 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3781 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3841 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3901 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3961 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4021 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4081 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4141 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     4201 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     4261 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     4321 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     4381 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     4441 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     4501 aaaacgacgg ccagtcttaa gctcgggccc
//
