LOCUS       sequence-335549-        4677 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 4677)
  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 4677)
  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-SLC15A1_STOP
FEATURES             Location/Qualifiers
     source          1..4677
                     /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(2234..2333)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2350..2369)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2351..2369)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2374..2390)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2374..2390)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2492..2511)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2503..3312
                     /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(2570..2589)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3459..4047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3948..3967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4201..4218
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4377..4404)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4496..4582)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4631..4653
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4645..4662
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4646..4662
                     /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 ccaccatggg catgtctaag
      121 agccactcct tctttggcta ccctctgtct atcttcttta tcgtggtgaa cgagttctgc
      181 gagcggttca gctactatgg catgagggcc atcctgatcc tgtatttcac caactttatc
      241 agctgggacg ataatctgtc caccgccatc taccacacct tcgtggccct gtgctatctg
      301 accccaatcc tgggcgccct gatcgcagac agctggctgg gcaagtttaa gacaatcgtg
      361 tctctgagca tcgtgtacac catcggccag gccgtgacat ccgtgagctc catcaacgac
      421 ctgaccgatc acaatcacga cggcacacca gattccctgc ccgtgcacgt ggtgctgtct
      481 ctgatcggac tggccctgat cgccctggga accggaggca tcaagccatg cgtgtccgcc
      541 ttcggcggcg atcagtttga ggagggccag gagaagcagc ggaacagatt cttttctatc
      601 ttctatctgg ccatcaatgc cggcagcctg ctgtccacca tcatcacacc catgctgagg
      661 gtgcagcagt gcggcatcca ctccaagcag gcctgttacc ctctggcctt cggcgtgcca
      721 gccgccctga tggcagtggc cctgatcgtg ttcgtgctgg gctctggcat gtataagaag
      781 tttaagcccc agggcaacat catgggcaag gtggccaagt gtatcggctt cgccatcaag
      841 aataggtttc gccaccggtc taaggccttc cctaagagag agcactggct ggactgggcc
      901 aaggagaagt acgatgagcg cctgatcagc cagatcaaga tggtgaccag agtgatgttt
      961 ctgtatatcc cactgcccat gttctgggcc ctgtttgacc agcagggaag caggtggacc
     1021 ctgcaggcaa ccacaatgtc cggcaagatc ggcgccctgg agatccagcc cgaccagatg
     1081 cagacagtga acgccatcct gatcgtgatc atggtgccca tcttcgatgc cgtgctgtac
     1141 cctctgatcg ccaagtgcgg cttcaacttc acctccctga agaagatggc cgtgggcatg
     1201 gtgctggcct ctatggcctt cgtggtggcc gccatcgtgc aggtggagat cgataagaca
     1261 ctgcccgtgt tcccaaaggg caacgaggtg cagatcaagg tgctgaacat cggcaacaat
     1321 accatgaata tcagcctgcc aggcgagatg gtgacactgg gccccatgtc ccagaccaat
     1381 gccttcatga catttgacgt gaacaagctg accagaatca atatctctag cccaggatcc
     1441 cctgtgaccg cagtgacaga cgatttcaag cagggccaga ggcacacact gctggtgtgg
     1501 gcccctaacc actaccaggt ggtgaaggat ggcctgaacc agaagccaga gaagggcgag
     1561 aatggcatcc gcttcgtgaa cacctttaat gagctgatca ccatcacaat gtccggcaag
     1621 gtgtatgcca acatctcctc ttacaatgcc tctacatatc agttctttcc cagcggcatc
     1681 aagggcttca ccatcagctc cacagagatc ccccctcagt gccagcctaa cttcaatacc
     1741 ttttacctgg agtttggcag cgcctacaca tatatcgtgc agcggaagaa tgactcttgt
     1801 ccagaggtga aggtgttcga ggatatcagc gccaacaccg tgaatatggc cctgcagatc
     1861 ccccagtact ttctgctgac ctgcggcgag gtggtgttct ccgtgacagg cctggagttt
     1921 tcctattctc aggccccttc taacatgaag agcgtgctgc aggcaggatg gctgctgacc
     1981 gtggcagtgg gcaatatcat cgtgctgatc gtggcaggag caggacagtt cagcaagcag
     2041 tgggccgagt acatcctgtt tgccgccctg ctgctggtgg tgtgcgtgat cttcgccatc
     2101 atggcccgct tttacacata tatcaaccct gccgagatcg aggcccagtt cgacgaggat
     2161 gagaagaaga accggctgga gaagagcaat ccatacttca tgtccggcgc caattctcag
     2221 aagcagatgt aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     2281 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     2341 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     2401 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2461 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     2521 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2581 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2641 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2701 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2761 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2821 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2881 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2941 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3001 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     3061 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3121 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3181 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3241 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3301 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     3361 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     3421 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3481 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3541 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3601 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3661 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3721 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3781 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3841 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3901 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3961 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4021 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4081 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4141 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4201 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4261 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4321 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     4381 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     4441 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     4501 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     4561 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     4621 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
