LOCUS       sequence-262917-        5904 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 5904)
  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 5904)
  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_SLC4A10
FEATURES             Location/Qualifiers
     source          1..5904
                     /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(4029..4128)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4145..4164)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4146..4164)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4169..4185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4169..4185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4287..4306)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4298..5107
                     /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(4365..4384)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5254..5842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5743..5762
                     /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 accatggaga tcaaggatca gggcgcccag atggagcctc tgctgccaac
      721 ccggaacgat gaggaggccg tggtggacag gggaggaacc agatccatcc tgaagacaca
      781 cttcgagaag gaggacctgg agggccaccg gacactgttt atcggagtgc acgtgccact
      841 gggaggaaga aagagccacc ggagacacag gcacagggga cacaagcacc ggaagagaga
      901 tagggagagg gactctggac tggaggatgg aagggagagc ccttcctttg acaccccaag
      961 ccagagagtg cagttcatcc tgggcacaga ggacgatgac gaggagcaca tcccccacga
     1021 tctgttcacc gagctggacg agatctgctg gagagaagga gaggacgcag agtggaggga
     1081 gacagcaagg tggctgaagt ttgaggagga tgtggaggac ggaggagagc ggtggtccaa
     1141 gccttacgtg gccaccctgt ccctgcactc tctgttcgag ctgagatctt gtatcctgaa
     1201 cggcacagtg ctgctggata tgcacgccaa taccctggag gagatcgccg atatggtgct
     1261 ggaccagcag gtgagctccg gccagctgaa tgaggacgtg cggcacagag tgcacgaggc
     1321 cctgatgaag cagcaccacc accagaacca gaagaagctg accaatcgga tcccaatcgt
     1381 gagatccttc gccgatatcg gcaagaagca gtctgagccc aacagcatgg acaagaatgc
     1441 aggacaggtg gtgagcccac agtccgcccc tgcctgcgtg gagaacaaga atgacgtgag
     1501 cagggagaac tctaccgtgg actttagcaa gggactggga ggacagcaga agggacacac
     1561 atccccctgt ggcatgaagc agaggcacga gaagggacca cctcaccagc aggagaggga
     1621 ggtggatctg cacttcatga agaagatccc acccggcgcc gaggcctcca atatcctggt
     1681 gggcgagctg gagtttctgg accggaccgt ggtggccttc gtgagactgt ctccagccgt
     1741 gctgctgcag ggactggcag aggtgccaat cccaacacgg ttcctgttta tcctgctggg
     1801 acctctgggc aagggacagc agtaccacga gatcggcaga tccatcgcca ccctgatgac
     1861 agatgaggtg ttccacgacg tggcctataa ggccaaggat aggaacgacc tggtgagcgg
     1921 catcgatgag tttctggacc aggtgaccgt gctgcctcca ggcgagtggg atccctctat
     1981 caggatcgag ccccctaaga acgtgcctag ccaggagaag cgcaagatcc ccgccgtgcc
     2041 taatggaacc gcagcacacg gagaggcaga gccacacggc ggccacagcg gacctgagct
     2101 gcagaggaca ggacgcatct ttggcggcct gatcctggac atcaagcgga aggccccata
     2161 cttctggagc gacttccggg acgccttctc cctgcagtgc ctggcctctt tcctgtttct
     2221 gtattgcgcc tgtatgtccc ccgtgatcac attcggcgga ctgctgggag aggcaaccga
     2281 gggaaggatc agcgccatcg agtccctgtt cggcgcctct atgaccggca tcgcctacag
     2341 cctgtttgga ggacagccac tgaccatcct gggatccaca ggacccgtgc tggtgttcga
     2401 gaagatcctg ttcaagtttt gcaaggagta cggcctgtct tatctgagcc tgagggcaag
     2461 catcggactg tggaccgcca cactgtgcat catcctggtg gccacagacg cctctagcct
     2521 ggtgtgctac atcacccgct ttacagagga ggccttcgcc agcctgatct gtatcatctt
     2581 tatctacgag gccctggaga agctgttcga gctgtccgag gcctatccaa tcaacatgca
     2641 caatgatctg gagctgctga cccagtattc ttgcaactgc gtggagccac acaaccccag
     2701 caatggcaca ctgaaggagt ggcgggagtc caacatctct gccagcgaca tcatctggga
     2761 gaatctgacc gtgtctgagt gcaagagcct gcacggcgag tacgtgggca gagcctgtgg
     2821 ccacgatcac ccttatgtgc cagacgtgct gttctggtct gtgatcctgt tctttagcac
     2881 cgtgacactg tccgccaccc tgaagcagtt taagacaagc aggtacttcc ccaccaaggt
     2941 gcgctccatc gtgtctgatt tcgccgtgtt tctgaccatc ctgtgcatgg tgctgatcga
     3001 ctatgccatc ggcatccctt ccccaaagct gcaggtgccc agcgtgttca agcctacacg
     3061 ggatgacaga ggctggtttg tgacccctct gggccccaac ccttggtgga cagtgatcgc
     3121 cgccatcatc ccagccctgc tgtgcaccat cctgatcttt atggatcagc agatcacagc
     3181 cgtgatcatc aatagaaagg agcacaagct gaagaagggc tgcggctatc acctggacct
     3241 gctgatggtg gccgtgatgc tgggcgtgtg ctccatcatg ggactgcctt ggttcgtggc
     3301 agcaaccgtg ctgagcatca cacacgtgaa ctccctgaag ctggagagcg agtgctccgc
     3361 cccaggagag cagccaaagt tcctgggaat cagggagcag agggtgaccg gcctgatgat
     3421 ctttatcctg atgggctcca gcgtgttcat gaccagcatc ctgaagttca tcccaatgcc
     3481 cgtgctgtac ggcgtgtttc tgtatatggg cgccagctcc ctgaagggca tccagttctt
     3541 tgataggatc aagctgttct ggatgccagc caagcaccag cccgacttta tctacctgag
     3601 gcacgtgccc ctgcgcaagg tgcacctgtt caccatcatc cagatgtcct gtctgggcct
     3661 gctgtggatc atcaaggtgt ctcgcgccgc catcgtgttt cctatgatgg tgctggccct
     3721 ggtgttcgtg aggaagctga tggatctgct gtttaccaag cgcgagctga gctggctgga
     3781 tgacctgatg ccagagagca agaagaagaa gctggaggat gccgagaagg aggaggagca
     3841 gtctatgctg gccatggagg acgagggcac agtgcagctg cctctggagg gccactatag
     3901 ggatgaccca agcgtgatca atatctccga cgagatgtct aagaccgccc tgtggcgcaa
     3961 cctgctgatc acagccgata atagcaagga caaggagtct agcttccctt ctaagtcctc
     4021 tccaagcgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     4081 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     4141 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     4201 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     4261 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     4321 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     4381 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     4441 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     4501 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     4561 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     4621 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     4681 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     4741 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     4801 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     4861 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     4921 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     4981 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     5041 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     5101 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     5161 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     5221 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5281 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5341 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5401 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5461 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5521 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5581 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5641 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5701 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5761 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5821 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5881 ttgctggcct tttgctcaca tgtt
//
