LOCUS       sequence-335556-        4170 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 4170)
  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 4170)
  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-SLC16A2_STOP
FEATURES             Location/Qualifiers
     source          1..4170
                     /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(1727..1826)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1843..1862)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1844..1862)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1867..1883)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1867..1883)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1985..2004)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1996..2805
                     /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(2063..2082)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2952..3540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3441..3460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3694..3711
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3870..3897)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3989..4075)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4124..4146
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4138..4155
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4139..4155
                     /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 actgcagtct
      121 caggcaagcg aggaggcaaa gggaccatgg caggaggcag accaggagca gcaggagcct
      181 gtgggaagcc cagagcctga gtccgagcca gagccagagc ctgagccaga gccagtgcct
      241 gtgccacctc cagagccaca gccagagcct cagccactgc cagatcctgc accactgcca
      301 gagctggagt tcgagtctga gagggtgcac gagcctgagc caaccccaac agtggagacc
      361 aggggaacag caaggggatt tcagccacct gagggaggat tcggatgggt ggtggtgttt
      421 gccgccacct ggtgcaacgg ctccatcttc ggcatccaca attctgtggg catcctgtac
      481 agcatgctgc tggaggagga gaaggagaag aaccgccagg tggagtttca ggcagcatgg
      541 gtgggcgcac ttgctatggg tatgatattt ttctgctccc ctatcgtgtc tatcttcacc
      601 gaccggctgg gctgtagaat caccgcaaca gcaggagcag cagtggcctt catcggcctg
      661 cacaccagct cctttacatc tagcctgagc ctgcggtact tcacatatgg catcctgttt
      721 ggctgcggct gttccttcgc ctttcagcct tctctggtca tcctgggcca ctacttccag
      781 cggagactgg gactggcaaa cggagtggtg tccgccggct cctctatctt ttccatgtct
      841 ttcccatttc tgatccgcat gctgggcgac aagatcaagc tggcccagac ctttcaggtg
      901 ctgtccacat tcatgtttgt gctgatgctg ctgtctctga cctatcggcc cctgctgcct
      961 agctcccagg ataccccaag caagcggggc gtgagaacac tgcaccagag gttcctggcc
     1021 cagctgcgca agtacttcaa tatgcgggtg tttaggcagc gcacatatag aatctgggcc
     1081 ttcggcatcg cagcagccgc cctgggatac tttgtgccct atgtgcacct gatgaagtac
     1141 gtggaggagg agttctccga gatcaaggag acctgggtgc tgctggtgtg catcggagcc
     1201 acaagcggac tgggcagact ggtgtccgga cacatcagcg actccatccc aggcctgaag
     1261 aagatctatc tgcaggtgct gtcttttctg ctgctgggcc tgatgagcat gatgatcccc
     1321 ctgtgcaggg acttcggcgg cctgatcgtg gtgtgcctgt ttctgggcct gtgcgatggc
     1381 ttctttatca ccatcatggc accaatcgcc ttcgagctgg tgggaccaat gcaggcaagc
     1441 caggccatcg gctatctgct gggcatgatg gccctgccca tgatcgcagg accaccaatc
     1501 gcaggactgc tgaggaattg ttttggcgat taccacgtgg ccttctattt tgcaggagtg
     1561 cctccaatca tcggagccgt gatcctgttc tttgtgcctc tgatgcacca gagaatgttc
     1621 aagaaggagc agagggactc tagcaaggat aagatgctgg cccctgaccc agatccaaac
     1681 ggagagctgc tgcctggaag ccctaatcca gaggagccaa tctaagaccc agctttcttg
     1741 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1801 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     1861 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     1921 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1981 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2041 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2101 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2161 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2221 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2281 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2341 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     2401 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     2461 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2521 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2581 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     2641 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     2701 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     2761 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     2821 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     2881 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     2941 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3001 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3061 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3121 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3181 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3241 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3301 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3361 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3421 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3481 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3541 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3601 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3661 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3721 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3781 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     3841 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     3901 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     3961 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     4021 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     4081 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     4141 aaaacgacgg ccagtcttaa gctcgggccc
//
