LOCUS       sequence-263276-        6273 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 6273)
  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 6273)
  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
FEATURES             Location/Qualifiers
     source          1..6273
                     /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(4398..4497)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4514..4533)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4515..4533)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4538..4554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4538..4554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4656..4675)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4667..5476
                     /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(4734..4753)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5623..6211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6112..6131
                     /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 accatgagct ccgccccacg gagacccgca aagggagcag attctttctg
      721 cacaccagag ccagagagcc tgggacctgg aaccccagga tttccagaac aggaggagga
      781 cgagctgcac cgcaccctgg gagtggagcg gttcgaggag atcctgcagg aggcaggaag
      841 cagaggagga gaggagccag gcaggtctta cggagaggag gattttgagt atcacagaca
      901 gtctagccac cacatccacc acccactgag cacacacctg ccacctgacg caaggcgccg
      961 gaagacccct cagggcccag gaaggaagcc aagaaggcgc cccggagcat ctcctaccgg
     1021 agagacccca acaatcgagg agggagagga ggacgaggat gaggcaagcg aggcagaggg
     1081 agcaagagcc ctgacccagc cttccccagt gtctacacct tcctctgtgc agttctttct
     1141 gcaggaggac gatagcgccg acagaaaggc cgagaggaca tccccaagct ccccagcacc
     1201 tctgccacac caggaggcca cacctagggc cagcaagggc gcccaggccg gcacccaggt
     1261 ggaggaggcc gaggccgagg cagtggcagt ggcaagcgga accgcaggcg gcgacgatgg
     1321 cggcgcctcc ggcagacccc tgcctaaggc acagcctgga cacaggtctt acaacctgca
     1381 ggagcggaga aggatcggaa gcatgacagg agcagagcag gccctgctgc caagagtgcc
     1441 taccgatgag atcgaggcac agaccctggc aacagcagac ctggatctga tgaagtccca
     1501 cagattcgag gacgtgccag gcgtgcgccg gcacctggtg aggaagaatg ccaagggcag
     1561 cacacagtcc ggcagagagg gcagggagcc aggaccaacc cctagggcaa ggccaagggc
     1621 accccacaag cctcacgagg tgttcgtgga gctgaacgag ctgctgctgg ataagaatca
     1681 ggagcctcag tggagggaga cagcaaggtg gatcaagttc gaggaggacg tggaggagga
     1741 gaccgagcgg tggggcaagc cacacgtggc ctctctgagc tttcggagcc tgctggagct
     1801 gagaaggaca ctggcacacg gagccgtgct gctggacctg gatcagcaga ccctgccagg
     1861 agtggcacac caggtggtgg agcagatggt catctccgat cagatcaagg cagaggacag
     1921 ggcaaacgtg ctgcgggccc tgctgctgaa gcactctcac cccagcgacg agaaggattt
     1981 ctcttttcct cggaatatct ccgccggatc tctgggaagc ctgctgggac accaccacgg
     2041 acagggagcc gagagcgatc cacacgtgac agagccactg atgggaggag tgcctgagac
     2101 ccgcctggag gtggagagag agagggagct gccaccacct gcaccaccag caggcatcac
     2161 ccggtccaag tctaagcacg agctgaagct gctggagaag atccccgaga acgcagaggc
     2221 aacagtggtg ctggtgggat gcgtggagtt cctgagcagg cctaccatgg cctttgtgcg
     2281 cctgagggag gcagtggagc tggatgccgt gctggaggtg ccagtgcccg tgagattcct
     2341 gtttctgctg ctgggcccct ctagcgccaa catggattac cacgagatcg gcaggagcat
     2401 ctccacactg atgtccgaca agcagttcca cgaggccgcc tatctggcag atgagaggga
     2461 ggacctgctg accgccatca atgcctttct ggactgttcc gtggtgctgc ctccatctga
     2521 ggtgcaggga gaggagctgc tgcgcagcgt ggcacacttc cagcggcaga tgctgaagaa
     2581 gagagaggag cagggcaggc tgctgcctac aggagcagga ctggagccaa agtccgccca
     2641 ggataaggcc ctgctgcaga tggtggaggc agcaggagca gcagaggacg atccactgag
     2701 gaggaccggc agacctttcg gcggcctgat cagggacgtg agaaggcgct accctcacta
     2761 tctgagcgac tttagagatg cactggaccc tcagtgcctg gcagccgtga tcttcatcta
     2821 ctttgccgcc ctgtccccag ccatcacatt cggcggactg ctgggagaga agacccagga
     2881 cctgatcggc gtgagcgagc tgatcatgtc caccgccctg cagggcgtgg tgttttgcct
     2941 gctgggagca cagccactgc tggtcatcgg ctttagcgga ccactgctgg tgttcgagga
     3001 ggccttcttt tccttttgtt cctctaatca cctggagtat ctggtgggca gggtgtggat
     3061 cggcttctgg ctggtgtttc tggccctgct gatggtggcc ctggagggaa gcttcctggt
     3121 gagattcgtg agcaggttca cacaggagat cttcgccttt ctgatcagcc tgatctttat
     3181 ctacgagacc ttctataagc tggtgaagat cttccaggag caccctctgc acggctgttc
     3241 tgccagcaac agctccgagg tggatggagg agagaatatg acctgggcag gagcaaggcc
     3301 aacactggga ccaggaaacc ggtctctggc aggacagagc ggacagggca agcctagagg
     3361 acagccaaat acagccctgc tgtccctggt gctgatggca ggcaccttct ttatcgcctt
     3421 ctttctgagg aagttcaaga actctcgctt ctttccaggc cggatccgga gagtgatcgg
     3481 cgactttggc gtgcccatcg ccatcctgat catggtgctg gtggattact ccatcgagga
     3541 cacctataca cagaagctgt ccgtgccatc tggcttcagc gtgacagcac cagagaagag
     3601 gggatgggtc atcaaccctc tgggcgagaa gtctcctttt ccagtgtgga tgatggtggc
     3661 cagcctgctg ccagccatcc tggtgtttat cctgatcttc atggagaccc agatcaccac
     3721 actgatcatc tccaagaagg agcgcatgct gcagaagggc tctggcttcc acctggatct
     3781 gctgctgatc gtggccatgg gcggcatctg tgccctgttt ggactgccat ggctggcagc
     3841 agcaaccgtg cggtccgtga cacacgcaaa cgcactgaca gtgatgtcta aggccgtggc
     3901 ccccggcgat aagcctaaga tccaggaggt gaaggagcag cgcgtgaccg gactgctggt
     3961 ggccctgctg gtgggactgt ccatcgtgat cggcgacctg ctgaggcaga tcccactggc
     4021 cgtgctgttc ggcatctttc tgtacatggg cgtgacctct ctgaatggca tccagttcta
     4081 tgagcggctg cacctgctgc tgatgccccc taagcaccac cccgacgtga cctacgtgaa
     4141 gaaggtgcgc acactgcgga tgcacctgtt taccgccctg cagctgctgt gcctggccct
     4201 gctgtgggcc gtgatgtcca cagccgcctc tctggccttc ccttttatcc tgatcctgac
     4261 cgtgccactg cgcatggtgg tgctgaccag aatcttcaca gatagggaga tgaagtgtct
     4321 ggacgccaac gaggccgagc ccgtgtttga tgagcgggag ggcgtggacg agtataatga
     4381 gatgcccatg cctgtggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     4441 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     4501 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     4561 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     4621 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     4681 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     4741 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     4801 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     4861 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     4921 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     4981 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     5041 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     5101 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     5161 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     5221 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     5281 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     5341 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     5401 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     5461 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     5521 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     5581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6241 cctggccttt tgctggcctt ttgctcacat gtt
//
