LOCUS       sequence-263659-        5787 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 5787)
  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 5787)
  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_SLC4A4
FEATURES             Location/Qualifiers
     source          1..5787
                     /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(3912..4011)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4028..4047)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4029..4047)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4052..4068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4052..4068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4170..4189)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4181..4990
                     /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(4248..4267)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5137..5725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5626..5645
                     /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 accatggagg acgaggccgt gctggatagg ggagcaagct tcctgaagca
      721 cgtgtgcgac gaggaggagg tggagggcca ccacaccatc tacatcggcg tgcacgtgcc
      781 caagagctat cggagaaggc gccggcacaa gagaaagaca ggccacaagg agaagaagga
      841 gaaggagagg atctccgaga actactccga caagtctgat atcgagaatg ccgacgagag
      901 ctcctctagc atcctgaagc ccctgatctc tcctgcagca gagaggatcc ggttcatcct
      961 gggagaggag gacgatagcc cagcaccacc tcagctgttt accgagctgg acgagctgct
     1021 ggccgtggat ggccaggaga tggagtggaa ggagacagcc cgctggatca agttcgagga
     1081 gaaggtggag cagggaggag agcggtggtc taagcctcac gtggccaccc tgagcctgca
     1141 ctccctgttt gagctgcgca catgtatgga gaagggcagc atcatgctgg accgggaggc
     1201 ctcctctctg cctcagctgg tggagatgat cgtggaccac cagatcgaga ccggcctgct
     1261 gaagccagag ctgaaggata aggtgaccta tacactgctg cgcaagcacc ggcaccagac
     1321 aaagaagtcc aacctgagat ctctggccga catcggcaag accgtgagct ccgcctctag
     1381 gatgttcaca aaccccgata atggcagccc tgccatgacc caccgcaacc tgacatctag
     1441 ctccctgaat gacatctccg ataagccaga gaaggaccag ctgaagaaca agttcatgaa
     1501 gaagctgccc cgggatgccg aggcctctaa tgtgctggtg ggcgaggtgg actttctgga
     1561 taccccattc atcgcctttg tgagactgca gcaggccgtg atgctgggcg ccctgaccga
     1621 ggtgcctgtg ccaacaaggt tcctgtttat cctgctgggc cccaagggca aggccaagtc
     1681 ctaccacgag atcggcagag ccatcgccac cctgatgtct gacgaggtgt tccacgatat
     1741 cgcctataag gccaaggaca ggcacgatct gatcgccggc atcgacgagt ttctggatga
     1801 agtgatcgtg ctgccaccag gagagtggga cccagccatc agaatcgagc ctccaaagag
     1861 cctgccttct agcgataaga ggaagaacat gtactccggc ggcgagaacg tgcagatgaa
     1921 tggcgacacc cctcacgatg gaggacacgg aggaggagga cacggcgact gcgaggagct
     1981 gcagagaaca ggcaggttct gtggcggcct gatcaaggat atcaagagga aggccccatt
     2041 ctttgcctct gacttttacg atgccctgaa tatccaggcc ctgagcgcca tcctgttcat
     2101 ctatctggcc accgtgacaa acgccatcac ctttggcgga ctgctgggcg acgcaacaga
     2161 taatatgcag ggcgtgctgg agtccttcct gggaaccgcc gtgtctggag ccatcttctg
     2221 cctgtttgcc ggccagccac tgaccatcct gtcctctaca ggccccgtgc tggtgtttga
     2281 gcgcctgctg ttcaacttca gcaaggacaa caatttcgat tacctggagt ttcggctgtg
     2341 gatcggcctg tggagcgcct tcctgtgcct gatcctggtg gccaccgacg cctcctttct
     2401 ggtgcagtac ttcacccgct ttacagagga gggcttcagc tccctgatct ccttcatctt
     2461 tatctatgac gcctttaaga agatgatcaa gctggccgat tactatccca tcaactccaa
     2521 tttcaaggtc ggctacaaca ccctgttttc ttgcacatgc gtgccccctg accctgccaa
     2581 catcagcatc tccaatgata ccacactggc cccagagtac ctgcccacca tgtctagcac
     2641 agacatgtat cacaatacca cattcgattg ggcctttctg agcaagaagg agtgctccaa
     2701 gtatggcggc aacctggtgg gcaacaattg taatttcgtg cccgacatca ccctgatgtc
     2761 cttcatcctg tttctgggca cctacacatc ctctatggcc ctgaagaagt tcaagacatc
     2821 tccatatttt cccaccacag ccagaaagct gatcagcgat ttcgccatca tcctgtccat
     2881 cctgatcttt tgcgtgatcg acgccctggt gggcgtggat accccaaagc tgatcgtgcc
     2941 cagcgagttc aagcctacat ccccaaacag gggctggttc gtgccaccct ttggcgagaa
     3001 tccttggtgg gtgtgcctgg cagcagcaat cccagccctg ctggtgacca tcctgatctt
     3061 catggaccag cagatcacag ccgtgatcgt gaaccgcaag gagcacaagc tgaagaaggg
     3121 cgccggctac cacctggatc tgttttgggt ggccatcctg atggtcatct gcagcctgat
     3181 ggcactgcca tggtacgtgg cagcaaccgt gatctctatc gcccacatcg acagcctgaa
     3241 gatggagacc gagacatccg cccctggaga gcagccaaag ttcctgggcg tgcgcgagca
     3301 gagggtgacc ggcacactgg tgtttatcct gaccggcctg agcgtgttca tggccccaat
     3361 cctgaagttt atccccatgc ctgtgctgta cggcgtgttc ctgtatatgg gcgtggcctc
     3421 cctgaatggc gtgcagttta tggaccgcct gaagctgctg ctgatgcctc tgaagcacca
     3481 gccagatttc atctacctgc ggcacgtgcc tctgagaagg gtgcacctgt tcacctttct
     3541 gcaggtgctg tgcctggccc tgctgtggat cctgaagagc acagtggccg ccatcatctt
     3601 cccagtgatg atcctggccc tggtggccgt gcggaagggc atggactacc tgttctctca
     3661 gcacgatctg agctttctgg acgatgtgat ccccgagaag gacaagaaga agaaggagga
     3721 tgagaagaag aagaagaaga agaagggctc tctggacagc gataacgacg attccgactg
     3781 cccttattct gagaaggtgc ccagcatcaa gatccctatg gatatcatgg agcagcagcc
     3841 attcctgtct gacagcaagc cctccgatag agagaggtct cctacctttc tggagagaca
     3901 cacatcctgt gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     3961 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     4021 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     4081 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     4141 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     4201 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     4261 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     4321 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     4381 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     4441 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     4501 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     4561 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4621 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4681 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4741 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4801 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4861 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4921 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     4981 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     5041 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     5101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5281 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5761 cttttgctgg ccttttgctc acatgtt
//
