LOCUS       sequence-479331-        4092 bp    DNA     circular SYN 04-AUG-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 4092)
  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 4092)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 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-SLC41A1_STOP
FEATURES             Location/Qualifiers
     source          1..4092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..47)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(201..789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(281..300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(936..1745)
                     /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     1659..1678
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1737..1756
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1858..1874
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1858..1874
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1879..1898
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1879..1897
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1915..2014
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3563..3662)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3678..3695)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3678..3694)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3687..3709)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3758..3844
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3936..3963
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
       61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg
      841 taactcacgt taagggattt tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg
      901 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
      961 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1021 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1081 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1141 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     1201 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1261 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1321 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1381 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1441 tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1501 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1561 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     1621 tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     1681 tataaatcag catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg
     1741 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     1801 atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgccag
     1861 gaaacagcta tgaccatgta atacgactca ctatagggga tatcagctgg atggcaaata
     1921 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt
     1981 cttataatgc caactttgta caagaaagct gggtcttagt cgcccacgtc ggtatctctg
     2041 tcgccgatca gccacagcac gtgaaaagac agggccagca gtcctgttcc cagcagatcg
     2101 cccagggcgg tcagataggg gatggagaag ttatctgggt ccagtcccct tccccacatc
     2161 cagtgcacca tccagtcggc gatatacagc aggatcagca cctgcagcag ggcggctgtc
     2221 atgtaaaaga tgatgaagat cagggtcagt gtggtgtggc cgccctgcat acagctgatt
     2281 gtgtacagaa acaccaggtg gccaggcacc accagcagga acagcactct ggcgctccga
     2341 gaattcacgt cgggggaaaa gaatgtggta catgggctag ggcagcgcct aggtgcctgc
     2401 tcggaattct cgccgggcat gccgttcatg tgcagaaagg tgctgatcct agatgcctgc
     2461 actgccacca gatttcctcc cactccgttg atcactggtg taaacacggc catgccggcg
     2521 aagttagggt cgctcacggt cttatccagg atcagtcctc ccacggagct gattgccatt
     2581 gcgatgatca ctggctccca tccgctatac agcacctccc tggttgctgg agatctccgt
     2641 gccagcacca cccacactgg cagcagggcc acaaagaagg cgcacaccag aggatagatg
     2701 tacctccagt gattcagctc caggtacagt ccccaggaga ttccgctcag cagggccagt
     2761 gtgatcaggt cgcccaggct tgctgcgata ggggttgcca cattatcggg gttgatgccg
     2821 atcttccgag agccgatgat cacgccgatc atgatcatgc ccagcaccag ggaggcgatg
     2881 aaggctgtgg ccacagagga ggcgcacagc agaaaggcgt gtgggatgga gaagtggccg
     2941 tcagggatcc agccaaacac cacggctgcg atgcttgcca ggaagcccac cacggttgcc
     3001 tgcacctgga tcagggccat gttgcctgtg atcattctcc acagctcctt gggggtatcc
     3061 atgtggccga tattggctgc tgtagacagc ctggatgcca gggtcatctc caggttgccc
     3121 ttcaggccca gcagggctgg caccaggata aacacctctg tcaccttctg gaacacctcc
     3181 cagtgctgca cgatatccag caccattcct gctgccacgg ttccaaatcc tgccagcaga
     3241 aaagggaaca gcacctgcag gccgatggag aagcttgtct ccttcagtgg ggaaggtggt
     3301 gcaggtcccc tatcggtgct cacgtcgtcg gactcattag actgggagcc gttctccagc
     3361 agtgcgtcct cctcgcgcac gccctttgca tttgccctgc tctcgatgac cacctccact
     3421 cctgctccat ctggtcccag gaactcagat gtgcctgcca gtggctccct tcctggtccg
     3481 tcgctagagc aagggctggc agatggtccg gttccgttca gctggtgcac atccttgggc
     3541 tctggcttgg agctcatggt ggagcctgct tttttgtaca aagttggcat tataaaaaag
     3601 cattgctcat caatgtgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     3661 tggggcccga gcttaagact ggccgtcgtt ttacaacgtc gtgactggga aaacatccat
     3721 gctagcgtta acgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     3781 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     3841 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
     3901 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
     3961 ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg tattaattgc gttgcgctca
     4021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4081 gcggggagag gc
//
