LOCUS       sequence-338291-        6276 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 6276)
  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 6276)
  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-SLC4A2_STOP
FEATURES             Location/Qualifiers
     source          1..6276
                     /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(3833..3932)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3949..3968)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3950..3968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3973..3989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3973..3989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4091..4110)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4102..4911
                     /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(4169..4188)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5058..5646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5547..5566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5800..5817
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5976..6003)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6095..6181)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6230..6252
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6244..6261
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6245..6261
                     /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 ccaccatgag ctccgcccca
      121 cggagacccg caaagggagc agattctttc tgcacaccag agccagagag cctgggacct
      181 ggaaccccag gatttccaga acaggaggag gacgagctgc accgcaccct gggagtggag
      241 cggttcgagg agatcctgca ggaggcagga agcagaggag gagaggagcc aggcaggtct
      301 tacggagagg aggattttga gtatcacaga cagtctagcc accacatcca ccacccactg
      361 agcacacacc tgccacctga cgcaaggcgc cggaagaccc ctcagggccc aggaaggaag
      421 ccaagaaggc gccccggagc atctcctacc ggagagaccc caacaatcga ggagggagag
      481 gaggacgagg atgaggcaag cgaggcagag ggagcaagag ccctgaccca gccttcccca
      541 gtgtctacac cttcctctgt gcagttcttt ctgcaggagg acgatagcgc cgacagaaag
      601 gccgagagga catccccaag ctccccagca cctctgccac accaggaggc cacacctagg
      661 gccagcaagg gcgcccaggc cggcacccag gtggaggagg ccgaggccga ggcagtggca
      721 gtggcaagcg gaaccgcagg cggcgacgat ggcggcgcct ccggcagacc cctgcctaag
      781 gcacagcctg gacacaggtc ttacaacctg caggagcgga gaaggatcgg aagcatgaca
      841 ggagcagagc aggccctgct gccaagagtg cctaccgatg agatcgaggc acagaccctg
      901 gcaacagcag acctggatct gatgaagtcc cacagattcg aggacgtgcc aggcgtgcgc
      961 cggcacctgg tgaggaagaa tgccaagggc agcacacagt ccggcagaga gggcagggag
     1021 ccaggaccaa cccctagggc aaggccaagg gcaccccaca agcctcacga ggtgttcgtg
     1081 gagctgaacg agctgctgct ggataagaat caggagcctc agtggaggga gacagcaagg
     1141 tggatcaagt tcgaggagga cgtggaggag gagaccgagc ggtggggcaa gccacacgtg
     1201 gcctctctga gctttcggag cctgctggag ctgagaagga cactggcaca cggagccgtg
     1261 ctgctggacc tggatcagca gaccctgcca ggagtggcac accaggtggt ggagcagatg
     1321 gtcatctccg atcagatcaa ggcagaggac agggcaaacg tgctgcgggc cctgctgctg
     1381 aagcactctc accccagcga cgagaaggat ttctcttttc ctcggaatat ctccgccgga
     1441 tctctgggaa gcctgctggg acaccaccac ggacagggag ccgagagcga tccacacgtg
     1501 acagagccac tgatgggagg agtgcctgag acccgcctgg aggtggagag agagagggag
     1561 ctgccaccac ctgcaccacc agcaggcatc acccggtcca agtctaagca cgagctgaag
     1621 ctgctggaga agatccccga gaacgcagag gcaacagtgg tgctggtggg atgcgtggag
     1681 ttcctgagca ggcctaccat ggcctttgtg cgcctgaggg aggcagtgga gctggatgcc
     1741 gtgctggagg tgccagtgcc cgtgagattc ctgtttctgc tgctgggccc ctctagcgcc
     1801 aacatggatt accacgagat cggcaggagc atctccacac tgatgtccga caagcagttc
     1861 cacgaggccg cctatctggc agatgagagg gaggacctgc tgaccgccat caatgccttt
     1921 ctggactgtt ccgtggtgct gcctccatct gaggtgcagg gagaggagct gctgcgcagc
     1981 gtggcacact tccagcggca gatgctgaag aagagagagg agcagggcag gctgctgcct
     2041 acaggagcag gactggagcc aaagtccgcc caggataagg ccctgctgca gatggtggag
     2101 gcagcaggag cagcagagga cgatccactg aggaggaccg gcagaccttt cggcggcctg
     2161 atcagggacg tgagaaggcg ctaccctcac tatctgagcg actttagaga tgcactggac
     2221 cctcagtgcc tggcagccgt gatcttcatc tactttgccg ccctgtcccc agccatcaca
     2281 ttcggcggac tgctgggaga gaagacccag gacctgatcg gcgtgagcga gctgatcatg
     2341 tccaccgccc tgcagggcgt ggtgttttgc ctgctgggag cacagccact gctggtcatc
     2401 ggctttagcg gaccactgct ggtgttcgag gaggccttct tttccttttg ttcctctaat
     2461 cacctggagt atctggtggg cagggtgtgg atcggcttct ggctggtgtt tctggccctg
     2521 ctgatggtgg ccctggaggg aagcttcctg gtgagattcg tgagcaggtt cacacaggag
     2581 atcttcgcct ttctgatcag cctgatcttt atctacgaga ccttctataa gctggtgaag
     2641 atcttccagg agcaccctct gcacggctgt tctgccagca acagctccga ggtggatgga
     2701 ggagagaata tgacctgggc aggagcaagg ccaacactgg gaccaggaaa ccggtctctg
     2761 gcaggacaga gcggacaggg caagcctaga ggacagccaa atacagccct gctgtccctg
     2821 gtgctgatgg caggcacctt ctttatcgcc ttctttctga ggaagttcaa gaactctcgc
     2881 ttctttccag gccggatccg gagagtgatc ggcgactttg gcgtgcccat cgccatcctg
     2941 atcatggtgc tggtggatta ctccatcgag gacacctata cacagaagct gtccgtgcca
     3001 tctggcttca gcgtgacagc accagagaag aggggatggg tcatcaaccc tctgggcgag
     3061 aagtctcctt ttccagtgtg gatgatggtg gccagcctgc tgccagccat cctggtgttt
     3121 atcctgatct tcatggagac ccagatcacc acactgatca tctccaagaa ggagcgcatg
     3181 ctgcagaagg gctctggctt ccacctggat ctgctgctga tcgtggccat gggcggcatc
     3241 tgtgccctgt ttggactgcc atggctggca gcagcaaccg tgcggtccgt gacacacgca
     3301 aacgcactga cagtgatgtc taaggccgtg gcccccggcg ataagcctaa gatccaggag
     3361 gtgaaggagc agcgcgtgac cggactgctg gtggccctgc tggtgggact gtccatcgtg
     3421 atcggcgacc tgctgaggca gatcccactg gccgtgctgt tcggcatctt tctgtacatg
     3481 ggcgtgacct ctctgaatgg catccagttc tatgagcggc tgcacctgct gctgatgccc
     3541 cctaagcacc accccgacgt gacctacgtg aagaaggtgc gcacactgcg gatgcacctg
     3601 tttaccgccc tgcagctgct gtgcctggcc ctgctgtggg ccgtgatgtc cacagccgcc
     3661 tctctggcct tcccttttat cctgatcctg accgtgccac tgcgcatggt ggtgctgacc
     3721 agaatcttca cagataggga gatgaagtgt ctggacgcca acgaggccga gcccgtgttt
     3781 gatgagcggg agggcgtgga cgagtataat gagatgccca tgcctgtgta agacccagct
     3841 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     3901 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     3961 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     4021 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     4081 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     4141 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     4201 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     4261 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     4321 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     4381 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     4441 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     4501 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     4561 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     4621 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     4681 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     4741 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     4801 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     4861 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     4921 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     4981 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     5041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5701 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5761 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5821 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5881 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     5941 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     6001 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     6061 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     6121 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     6181 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     6241 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
