LOCUS       sequence-479326-        4989 bp    DNA     circular SYN 26-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 4989)
  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 4989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 26, 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-SLC9A2_STOP
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(66..165)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(181..198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(181..197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(190..212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      261..347
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      439..466
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(625..642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(796..1384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(876..895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1531..2340)
                     /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     2254..2273
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2332..2351
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2453..2469
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2453..2469
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2474..2493
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2474..2492
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2510..2609
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cagtaataac ataggtggca gtggtgccct cagggagcgc cagttgccca gaggctccat
       61 ggtggagcct gcttttttgt acaaagttgg cattataaaa aagcattgct catcaatgtg
      121 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggggcc cgagcttaag
      181 actggccgtc gttttacaac gtcgtgactg ggaaaacatc catgctagcg ttaacgcgag
      241 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac tgggcctttc
      301 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
      361 atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
      421 ccaggcatca aactaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
      481 ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg ctttccagtc
      541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact
     1441 cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg
     1501 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     1561 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1621 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1681 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1741 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     1801 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1861 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1921 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1981 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc
     2041 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     2101 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     2161 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     2221 gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     2281 atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat
     2341 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     2401 tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac
     2461 agctatgacc atgtaatacg actcactata ggggatatca gctggatggc aaataatgat
     2521 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gctttcttat
     2581 aatgccaact ttgtacaaga aagctgggtc ttagggcttc tcgcttccga accttgcctt
     2641 cctagatcct ggctcggatg ccctccacac cagccttgga ggtggctttg gagggatgcc
     2701 ctcggtcaga gagtcctctc tctcagagcc ggactgatcc ttggacagca gaggctgctg
     2761 cagcaggcca gatgtctggg tgcccttctc cctggagtga ggtgtgggtg gggtagaggg
     2821 catgtctctg ccggagtcca catccacctc attcttccac tccatcttat aggactgggg
     2881 gctcttctta gaaaactgct cgggcaggaa ccttctggcc cttggctgca gattcagcac
     2941 tgtggttcct gcgtctgcat cggagtcgct agagttgcca tcggcgatgc tgatggtccg
     3001 gcgcttctgc agcttctctg gcagctttgt attcttaggc aggctcaggt atctgcttgt
     3061 agaggtggag gccctgtgct ctctgttcag ggagctgtcc ttcctgatgc tctctctcag
     3121 agagtgcctt ctccggatca ggatctcctt ggcctgccgc tcagatgtat cggcggtcag
     3181 ggagtggcga ttatagctca gggtcctctg tctgatctgg tacaggttcc tggacagcag
     3241 ctctctgatc tcgtctgtct cagaggaggt caccttccgg atcttctcct cgcgacaatc
     3301 gttcagggag gcgaatgttg gcacggtgct gatcatgcct gtctcggcca tctcgatggc
     3361 gtgcttgatc tccagcttct tatacaggga cacgatgcta gacttaggct gattctccct
     3421 gatcagcagc tttctcaggt acttatcgtc aaacttcttg aacttgtccc tccaaaagtt
     3481 gtggccccag tgtccgcaca cgtcctcgat tccggtcttc acgtgatcga acagtctgca
     3541 atagatctcc tcgctcacgg cctgctgctt cttattagac ctcttcacat ccagaaactc
     3601 caccaggggt ctgattgtga tgcccaggat gaacacggtg aagaagatga ccacgatggc
     3661 ggcggtgata aacagcttct tccgagggaa cacggctgct ggcagcagaa acaccagtgc
     3721 gaaacagatt gctcccctca gtcctccgta tgcgatgata aactggtcct tgaatgtcag
     3781 tgggatggtc cgaaagcggt tgatgacctg ggtcagcacg aacacgccca gggccctcca
     3841 catcagacag aaggccagtg taaagcacac gaaggcccaa ttccactcgt ggttcttgcc
     3901 cacggtgctc acgcccataa agatgaagat cagtgtctca gacacggagc tcagcatctt
     3961 catgaagtac ttgattgtgg tgtaggactt ctggctcaca ttctcctcca catacttgtt
     4021 cattgtcatg gcgcaggcgg tgatggccat gatgccggac aggtggaaca tctcggcggt
     4081 gatgtaagac agataggagt acagaaacac gaacaggggc tcgatcactc tgatattgtg
     4141 tgtaaagcgt gtggtgaagg cggcgataaa gcccaggaag attccgatca gcacgcctcc
     4201 gattcccacc acaaagaagt tggcgatgcc ggcaaacacg tcgattgtct cgatggtctt
     4261 catctgacaa aaggacttga acagattata cagcaccacg gtcacggcat cgttcagcag
     4321 gctctcgcca aacaccagga tgtacagctg ctcattcacg tggatgttct cgaacacggc
     4381 cagcacggcc acagggtcca cggcgctgat cagagagccg aacagcaggt tctgcagcag
     4441 ggtgatatcg ctcaggccaa aggcctctat ttgacaaatg ccgaacaggc tcacgccgat
     4501 gccgatagaa ttccacaggg tgcccaccac ggcgtaccaa aagattgtgc cgatgttctc
     4561 aaagaaaggg cgggtgggca tgaaatagcc tgcgtccagc acgataggtg gcagcaggta
     4621 cagaaagaac acgtcggtct tcatggctgg aggggacttc tcatccacgc cgaagatgat
     4681 gccgcccagc agcaggccca ccatgatcag caggcagctc tcgggcacga ttgttggcag
     4741 cttgtgatac aggtgaaagc cgatcttggc cagagaggcc agcaggatcc acagggtgat
     4801 ctcgaatggg atctgcacgt gagggtaatc cagtgtaaac acgggcagcc ggctctcctc
     4861 gaacagtgtg gttcctggtg ccaccacgga tgctgggctt ggtggagagg agctggtgcc
     4921 cattgcccta ggtgcattca gcagtgtctc ggccagggcg cccactggtc ctgccacttg
     4981 cagcagcag
//
