LOCUS       sequence-263159-        6246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6246)
  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 6246)
  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_SLC4A3
FEATURES             Location/Qualifiers
     source          1..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4371..4470)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4487..4506)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4488..4506)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4511..4527)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4511..4527)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4629..4648)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4640..5449
                     /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(4707..4726)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5596..6184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6085..6104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggcca acggcgtgat cccacctcca ggaggagcaa gcccactgcc
      721 tcaggtgcgg gtgcccctgg aggagccccc tctgtcccct gacgtggagg aggaggacga
      781 tgacctgggc aagaccctgg ccgtgtctag atttggcgat ctgatcagca agccaccagc
      841 atgggaccca gagaagcctt ctcggagcta ttccgagaga gatttcgagt ttcacaggca
      901 cacaagccac cacacccacc acccactgtc cgccaggctg cctccacccc acaagctgcg
      961 gagactgcct ccaacatccg ccaggcacac caggcgcaag cgcaagaagg agaagacatc
     1021 cgccccacct tctgagggaa ccccaccaat ccaggaggag ggaggagcag gcgtggacga
     1081 ggaagaagag gaagaagaag aggaggaggg agagagcgag gcagagcctg tggagcctcc
     1141 acactccgga accccacaga aggccaagtt ctctatcggc agcgacgagg atgacagccc
     1201 aggactgcca ggaagggcag cagtgacaaa gccactgccc tctgtgggcc cacacaccga
     1261 taagagcccc cagcacagct cctctagccc tagcccaagg gcaagggcat ccaggctggc
     1321 aggcgagaag tctagacctt ggtctccaag cgcctcctac gacctgagag agaggctgtg
     1381 cccaggaagc gccctgggaa acccaggagg ccctgagcag caggtgccta cagatgaggc
     1441 agaggcacag atgctgggat ccgccgacct ggatgacatg aagtctcaca ggctggagga
     1501 taatccaggc gtgcggagac acctggtgaa gaagccaagc aggacccagg gaggaagggg
     1561 atctccaagc ggactggcac caatcctgag gcggaagaag aagaagaaga agctggaccg
     1621 gagaccccac gaggtgttcg tggagctgaa cgagctgatg ctggatcgct ctcaggagcc
     1681 tcactggcgg gagacagcca gatggatcaa gtttgaggag gacgtggagg aggagaccga
     1741 gaggtggggc aagcctcacg tggcctccct gtctttccgc agcctgctgg agctgaggag
     1801 gaccatcgca cacggagccg ccctgctgga tctggagcag accacactgc caggcatcgc
     1861 ccacctggtg gtggagacaa tgatcgtgtc tgatcagatc aggcctgagg acagggcaag
     1921 cgtgctgagg accctgctgc tgaagcacag ccacccaaac gatgacaagg actccggctt
     1981 ctttccacgc aatccctcct ctagctccat gaactccgtg ctgggcaatc accaccccac
     2041 accttctcac ggacctgatg gagcagtgcc aaccatggca gatgacctgg gagagccagc
     2101 accactgtgg cctcacgatc cagacgcaaa ggagaagcca ctgcacatgc ctggaggcga
     2161 cggacacagg ggcaagtccc tgaagctgct ggagaagatc cccgaggatg cagaggcaac
     2221 agtggtgctg gtgggatgcg tgccctttct ggagcagcct gcagcagcct tcgtgcggct
     2281 gaatgaggcc gtgctgctgg agagcgtgct ggaggtgcct gtgccagtga gattcctgtt
     2341 tgtgatgctg ggcccttccc acacctctac agactatcac gagctgggcc ggtctatcgc
     2401 caccctgatg agcgataagc tgtttcacga ggccgcctac caggccgatg acagacagga
     2461 tctgctgagc gccatctccg agttcctgga cggctccatc gtgatcccac cttctgaggt
     2521 ggagggaagg gacctgctga gatccgtggc agcctttcag agggagctgc tgcgcaagcg
     2581 gagagagagg gagcagacaa aggtggagat gaccacaagg ggaggatata ccgcaccagg
     2641 caaggagctg agcctggagc tgggaggatc cgaggccaca ccagaggatg acccactgct
     2701 gaggaccgga tccgtgttcg gaggactggt gagagatgtg aggcgccggt acccccacta
     2761 tccttctgat ctgagggacg cactgcacag ccagtgcgtg gccgccgtgc tgttcatcta
     2821 ctttgccgcc ctgagccccg ccatcacatt tggcggactg ctgggagaga agaccgaggg
     2881 actgatgggc gtgtctgagc tgatcgtgag caccgccgtg ctgggcgtgc tgttctccct
     2941 gctgggagca cagccactgc tggtggtggg cttctctgga cctctgctgg tgtttgagga
     3001 ggccttcttt aagttctgta gggcccagga cctggagtat ctgacaggac gcgtgtgggt
     3061 gggactgtgg ctggtggtgt tcgtgctggc cctggtggca gcagagggat cctttctggt
     3121 gcggtacatc tctccattca cccaggagat cttcgccttt ctgatcagcc tgatcttcat
     3181 ctatgagacc ttctacaagc tgtataaggt gtttaccgag cacccactgc tgcccttcta
     3241 cccacccgag ggcgccctgg agggaagcct ggacgcaggc ctggagccta acggatccgc
     3301 cctgcctcca accgagggac ccccttctcc tagaaaccag ccaaatacag ccctgctgtc
     3361 cctgatcctg atgctgggca ccttctttat cgccttcttt ctgcgcaagt tccggaattc
     3421 tagatttctg ggcggcaagg ccagaaggat catcggcgac tttggcatcc ccatcagcat
     3481 cctggtcatg gtgctggtgg attactccat caccgacaca tatacccaga agctgacagt
     3541 gccaaccggc ctgtctgtga ccagccccga taagcggtcc tggtttatcc cacccctggg
     3601 ctctgccaga cctttccctc catggatgat ggtggcagca gcagtgccag ccctgctggt
     3661 gctgatcctg atcttcatgg agacacagat caccgccctg atcgtgtctc agaaggcacg
     3721 ccggctgctg aagggaagcg gctttcacct ggacctgctg ctgatcggaa gcctgggagg
     3781 actgtgcgga ctgttcggac tgccatggct gacagcagca accgtgcggt ccgtgacaca
     3841 cgtgaatgcc ctgacagtga tgagaaccgc catcgccccc ggcgataagc ctcagatcca
     3901 ggaggtgagg gagcagaggg tgaccggcgt gctgatcgca tccctggtgg gactgtctat
     3961 cgtgatgggc gccgtgctga gaaggatccc actggccgtg ctgttcggca tctttctgta
     4021 tatgggcgtg acaagcctgt ccggcatcca gctgtcccag aggctgctgc tgatcctgat
     4081 gcccgccaag caccaccctg agcagccata cgtgacaaag gtgaagacct ggcgcatgca
     4141 cctgttcacc tgcatccagc tgggctgtat cgccctgctg tgggtggtga agagcacagc
     4201 cgcctccctg gccttcccct ttctgctgct gctgaccgtg cctctgagac actgtctgct
     4261 gccacggctg tttcaggaca gagagctgca ggccctggat agcgaggacg cagagccaaa
     4321 cttcgatgag gacggccagg atgagtacaa tgagctgcac atgcccgtgg acccagcttt
     4381 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     4441 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     4501 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     4561 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     4621 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     4681 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     4741 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     4801 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     4861 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     4921 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     4981 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     5041 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     5101 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     5161 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     5221 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     5281 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     5341 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     5401 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     5461 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     5521 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     5581 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5641 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5701 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5761 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5821 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5881 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5941 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6001 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6061 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6121 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6181 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6241 catgtt
//
