LOCUS       sequence-420966-        5790 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 5790)
  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 5790)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..5790
                     /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(3915..4014)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4031..4050)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4032..4050)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4055..4071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4055..4071)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4173..4192)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4184..4993
                     /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(4251..4270)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5140..5728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5629..5648
                     /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 taagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     3961 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     4021 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     4081 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     4141 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     4201 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     4261 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     4321 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     4381 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     4441 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     4501 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4561 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4621 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4681 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     4741 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4801 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4861 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4921 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4981 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     5041 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     5101 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5161 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5221 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5281 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5341 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5401 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5461 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5521 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5581 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5641 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5701 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5761 ggccttttgc tggccttttg ctcacatgtt
//
