LOCUS       sequence-424385-        4503 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 4503)
  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 4503)
  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-SLC23A2_STOP
FEATURES             Location/Qualifiers
     source          1..4503
                     /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(2060..2159)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2176..2195)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2177..2195)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2200..2216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2200..2216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2318..2337)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2329..3138
                     /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(2396..2415)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3285..3873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3774..3793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4027..4044
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4203..4230)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4322..4408)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4457..4479
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4471..4488
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4472..4488
                     /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 ccaccatgat gggcatcggc
      121 aagaacacca caagcaagtc catggaggca ggaagctcca ccgagggcaa gtacgaggac
      181 gaggccaagc acccagcctt ctttacactg ccagtggtca tcaacggagg agccacctct
      241 agcggagagc aggacaatga ggatacagag ctgatggcca tctataccac agagaatggc
      301 atcgccgaga agtcctctct ggccgagaca ctggactcta ccggaagcct ggacccacag
      361 aggtccgata tgatctacac catcgaggat gtgccccctt ggtatctgtg catcttcctg
      421 ggcctgcagc actacctgac atgtttctct ggcaccatcg ccgtgccttt tctgctggcc
      481 gacgccatgt gcgtgggcta tgatcagtgg gccacaagcc agctgatcgg caccatcttc
      541 ttttgcgtgg gcatcaccac actgctgcag accacattcg gctgtcggct gcccctgttt
      601 caggcaagcg ccttcgcctt tctggcacct gccagagcca tcctgagcct ggacaagtgg
      661 aagtgtaaca ccacagacgt gagcgtggcc aatggcacag ccgagctgct gcacaccgag
      721 cacatctggt accctaggat ccgcgagatc cagggcgcca tcatcatgag ctccctgatc
      781 gaggtggtca tcggactgct gggactgcca ggcgccctgc tgaagtatat cggccccctg
      841 accatcacac ctaccgtggc cctgatcgga ctgtccggat tccaggcagc aggagagagg
      901 gcaggcaagc actggggcat cgccatgctg accatcttcc tggtgctgct gttttcccag
      961 tacgccagaa acgtgaagtt tccactgccc atctacaagt ctaagaaggg ctggacagcc
     1021 tataagctgc agctgttcaa gatgtttcca atcatcctgg ccatcctggt gagctggctg
     1081 ctgtgcttca tctttacagt gaccgacgtg ttcccacccg attccacaaa gtacggcttt
     1141 tatgccagga ccgatgcaag gcagggcgtg ctgctggtgg caccatggtt caaggtgcct
     1201 tatccatttc agtggggact gccaaccgtg tctgccgcag gcgtgatcgg aatgctgagc
     1261 gccgtggtgg ccagcatcat cgagtccatc ggcgactact atgcctgcgc aaggctgagc
     1321 tgtgcacctc caccacctat ccacgccatc aaccgcggca tcttcgtgga gggcctgtct
     1381 tgcgtgctgg atggcatctt tggcacaggc aacggcagca cctctagctc ccctaatatc
     1441 ggcgtgctgg gcatcaccaa agtgggctcc aggagagtga tccagtgcgg agccgccctg
     1501 atgctggccc tgggcatgat cggcaagttc tctgccctgt ttgcaagcct gccagaccct
     1561 gtgctgggcg ccctgttctg tacactgttt ggcatgatca ccgccgtggg cctgtctaat
     1621 ctgcagttca tcgatctgaa ctctagcagg aatctgttcg tgctgggctt ttccatcttc
     1681 tttggcctgg tgctgccttc ttacctgcgc cagaacccac tggtgacagg catcaccggc
     1741 atcgaccagg tgctgaatgt gctgctgacc acagccatgt tcgtgggcgg ctgcgtggcc
     1801 tttatcctgg ataacacaat cccaggaacc ccagaggaga ggggaatcag aaagtggaag
     1861 aagggcgtgg gcaagggcaa taagagcctg gacggcatgg agtcctacaa cctgcccttc
     1921 ggcatgaata tcatcaagaa gtacagatgt ttctcctatc tgcctatctc tccaacattt
     1981 gtgggctata cctggaaggg cctgcggaag tccgataatt ctagatcctc tgacgaggat
     2041 agccaggcca ccggctaaga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     2101 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     2161 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     2221 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     2281 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     2341 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     2401 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     2461 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     2521 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     2581 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     2641 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     2701 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     2761 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     2821 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     2881 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2941 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     3001 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     3061 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     3121 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     3181 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     3241 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3301 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3361 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3421 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3481 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3541 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3601 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3661 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3721 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3781 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3841 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3901 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3961 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4021 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4081 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4141 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     4201 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     4261 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     4321 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     4381 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     4441 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     4501 ccc
//
