LOCUS       sequence-479353-        5964 bp    DNA     circular SYN 23-JUL-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 5964)
  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 5964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 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-SLC4A5_STOP
FEATURES             Location/Qualifiers
     source          1..5964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     371..388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(547..574)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(666..752)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     801..823
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     815..832
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     816..832
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    848..947
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4368..4467)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4484..4503)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4485..4503)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4508..4524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4508..4524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4626..4645)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4637..5446
                     /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(4704..4723)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(5593..5964,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
      541 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
      601 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
      661 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
      721 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
      781 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
      841 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacacattga
      901 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgaaggt
      961 gaaggaggag aaggccggcg tgggcaagct ggaccacacc aaccaccgga gaaggttccc
     1021 cgatcagaag gagtgcccac ctatccacat cggactgcca gtgcctacct accctcagag
     1081 aaagacagat cagaagggcc acctgtctgg cctgcagaag gtgcactggg gactgaggcc
     1141 agaccagcca cagcaggagc tgaccggacc tggaagcgga gcaagctccc aggactctag
     1201 catggatctg atctctagaa caaggagccc agcagcagag cagctgcagg acatcctggg
     1261 agaggaggat gaggccccaa atcccaccct gtttacagag atggataccc tgcagcacga
     1321 cggcgatcag atggagtgga aggagagcgc ccgctggatc aagttcgagg agaaggtgga
     1381 ggagggagga gagcggtggt ccaagcccca cgtgagcacc ctgtccctgc actctctgtt
     1441 cgagctgaga acatgcctgc agaccggcac agtgctgctg gacctggatt ctggcagcct
     1501 gccacagatc atcgacgatg tgatcgagaa gcagatcgag gacggactgc tgaggccaga
     1561 gctgagggag cgggtgtctt acgtgctgct gcgccggcac aggcaccaga ccaagaagcc
     1621 tatccaccgg agcctggccg atatcggcaa gtccgtgtct accacaaaca gatcccctgc
     1681 aaggtctcca ggagcaggac caagcctgca ccactccaca gaggacctga gaatgaggca
     1741 gtccgccaac tatggccggc tgtgccacgc acagagccgg tccatgaatg acatctctct
     1801 gacccctaac acagatcagc ggaagaataa gtttatgaag aagatcccaa aggacagcga
     1861 ggcctccaat gtgctggtgg gcgaggtgga cttcctggat cagcccttca tcgcctttgt
     1921 gagactgatc cagtccgcca tgctgggagg agtgaccgag gtgcctgtgc caacaaggtt
     1981 cctgtttatc ctgctgggcc ctagcggcag agccaagtcc tacaacgaga tcggcagggc
     2041 catcgccacc ctgatggtgg acgatctgtt tagcgacgtg gcctataagg cccgcaatcg
     2101 ggaggatctg atcgccggca tcgacgagtt cctggatgaa gtgatcgtgc tgccaccagg
     2161 agagtgggac ccaaacatca gaatcgagcc tccaaagaag gtgcccagcg ccgataagag
     2221 gaagagcgtg ttcagcctgg ccgagctggg ccagatgaac ggctccgtgg gaggaggagg
     2281 aggagcacca ggaggaggca atggcggagg aggcggcggc ggctctggcg gcggcgccgg
     2341 cagcggcggc gccggcggca cctcctctgg cgacgatggc gagatgcccg ccatgcacga
     2401 gatcggcgag gagctgatct ggacaggccg cttctttggc ggcctgtgcc tggatatcaa
     2461 gcggaagctg ccttggttcc catccgactt ttacgatggc ttccacatcc agtctatcag
     2521 cgccatcctg tttatctatc tgggctgtat caccaacgcc atcacattcg gcggactgct
     2581 gggcgacgca accgataatt accagggcgt gatggagtcc tttctgggca cagccatggc
     2641 cggctccctg ttctgcctgt tttctggcca gcccctgatc atcctgagct ccaccggccc
     2701 tatcctgatc tttgagaagc tgctgttcga ctttagcaag ggcaacggcc tggattatat
     2761 ggagttccgc ctgtggatcg gactgcactc cgccgtgcag tgtctgatcc tggtggccac
     2821 cgacgcctct ttcatcatca agtacatcac ccggtttaca gaggagggct tctctacact
     2881 gatcagcttc atctttatct atgacgccat caagaagatg atcggcgcct ttaagtacta
     2941 tccaatcaac atggacttca agcccaattt catcaccaca tacaagtgcg agtgcgtggc
     3001 ccccgacacc gtgaacacca cagtgttcaa tgcaagcgcc ccactggcac ctgacaccaa
     3061 cgcctccctg tacaacctgc tgaatctgac agccctggat tggtccctgc tgtctaagaa
     3121 ggagtgcctg tcttatggcg gccgcctgct gggcaatagc tgtaagttta tccctgatct
     3181 ggccctgatg tccttcatcc tgttctttgg cacctactct atgaccctga cactgaagaa
     3241 gttcaagttt agcagatatt tcccaacaaa ggtgagggcc ctggtggccg acttttccat
     3301 cgtgttctct atcctgatgt tttgcggcat cgatgcctgt ttcggactgg agaccccaaa
     3361 gctgcacgtg cctagcgtga tcaagccaac aagacccgac aggggctggt ttgtggcccc
     3421 attcggcaag aacccctggt gggtgtatcc tgcctccatc ctgccagccc tgctggtgac
     3481 catcctgatc tttatggatc agcagatcac agccgtgatc gtgaacagaa aggagaataa
     3541 gctgaagaag gccgccggct accacctgga cctgttttgg gtgggcatcc tgatggccct
     3601 gtgcagcttc atgggcctgc cttggtacgt ggccgccacc gtgatcagca tcgcccacat
     3661 cgattccctg aagatggaga ccgagacatc cgcccctgga gagcagccac agttcctggg
     3721 cgtgcgcgag cagagggtga ccggcatcat cgtgtttatc ctgacaggca tcagcgtgtt
     3781 cctggcccca atcctgaagt gtatccccct gcctgtgctg tacggcgtgt ttctgtatat
     3841 gggcgtggcc tctctgaatg gaatccagat gggaaccgga ggaagcgagt tcaagatcca
     3901 gaagaagctg acaccctttt gggagcggtg caagctgttc ctgatgcccg ccaagcacca
     3961 gcctgaccac gcctttctgc ggcacgtgcc tctgagaagg atccacctgt tcaccctggt
     4021 gcagatcctg tgcctggccg tgctgtggat cctgaagtcc acagtggccg ccatcatctt
     4081 tcccgtgatg atcctgggcc tgatcatcgt gcgccggctg ctggatttca tcttttctca
     4141 gcacgacctg gcctggatcg ataacatcct gccagagaag gagaagaagg agaccgacaa
     4201 gaagagaaag aggaagaagg gagcacacga ggactgcgat gaggagcccc agttcccccc
     4261 tccaagcgtg atcaagatcc ctatggagag cgtgcagtcc gacccacaga atggcatcca
     4321 ctgtatcgcc cgcaagcggt ctagctcctg gtcctactct ctgtaagacc cagctttctt
     4381 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     4441 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     4501 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     4561 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     4621 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     4681 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     4741 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     4801 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     4861 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     4921 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     4981 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     5041 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     5101 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     5161 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     5221 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     5281 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     5341 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     5401 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     5461 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     5521 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     5581 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5641 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5701 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5761 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5821 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5881 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5941 ggagcgaacg acctacaccg aact
//
