LOCUS       sequence-312660-        4074 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 4074)
  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 4074)
  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-SLC7A5_STOP
FEATURES             Location/Qualifiers
     source          1..4074
                     /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"
     CDS             complement(1747..1758)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2199..2298)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2315..2334)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2316..2334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2339..2355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2339..2355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2457..2476)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2468..3277
                     /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(2535..2554)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3424..4012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3913..3932
                     /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 accatggccg gcgccggccc taagaggaga gccctggcag caccagcagc
      721 agaggagaag gaggaggcaa gggagaagat gctggcagca aagtccgccg acggatctgc
      781 cccagcagga gagggcgagg gcgtgaccct gcagcgcaac atcacactgc tgaatggcgt
      841 ggccatcatc gtgggcacca tcatcggctc cggcatcttc gtgaccccaa caggcgtgct
      901 gaaggaggcc ggctctccag gactggccct ggtggtgtgg gcagcatgcg gcgtgtttag
      961 catcgtgggc gccctgtgct acgcagagct gggcaccaca atcagcaagt ccggcggcga
     1021 ttacgcctat atgctggagg tgtatggcag cctgcccgcc ttcctgaagc tgtggatcga
     1081 gctgctgatc atccggccta gctcccagta catcgtggcc ctggtgttcg ccacctatct
     1141 gctgaagcct ctgtttccaa catgcccagt gcctgaggag gcagcaaagc tggtggcatg
     1201 cctgtgcgtg ctgctgctga ccgccgtgaa ctgttactcc gtgaaggccg ccacaagagt
     1261 gcaggacgcc ttcgccgcag caaagctgct ggccctggcc ctgatcatcc tgctgggctt
     1321 tgtgcagatc ggcaagggcg acgtgtctaa cctggatcct aatttcagct ttgagggcac
     1381 caagctggat gtgggcaaca tcgtgctggc cctgtactct ggcctgttcg cctacggcgg
     1441 ctggaactat ctgaattttg tgacagagga gatgatcaac ccctatagga atctgcctct
     1501 ggccatcatc atcagcctgc ctatcgtgac cctggtgtac gtgctgacaa atctggccta
     1561 tttcaccaca ctgtccaccg agcagatgct gtctagcgag gccgtggccg tggactttgg
     1621 caactaccac ctgggcgtga tgagctggat catcccagtg ttcgtgggcc tgagctgctt
     1681 tggctccgtg aatggctctc tgttcacctc ctctcggctg ttctttgtgg gctctagaga
     1741 gggccacctg ccctctatcc tgagcatgat ccaccctcag ctgctgacac cagtgcccag
     1801 cctggtgttc acctgcgtga tgacactgct gtatgccttc agcaaggata tcttttccgt
     1861 gatcaacttc ttttccttct ttaattggct gtgcgtggcc ctggccatca tcggcatgat
     1921 ctggctgagg cacaggaagc cagagctgga gaggcccatc aaggtgaacc tggccctgcc
     1981 agtgttcttt atcctggcct gcctgttcct gatcgccgtg tccttttgga agacccccgt
     2041 ggagtgtggc atcggcttca caatcatcct gtctggcctg cccgtgtact tcttcggcgt
     2101 gtggtggaag aataagccaa agtggctgct gcagggcatc ttttccacca ccgtgctgtg
     2161 ccagaagctg atgcaggtgg tgccccagga gacctaagac ccagctttct tgtacaaagt
     2221 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2281 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2341 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2401 agataaaata atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2461 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2521 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2581 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2641 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2701 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2761 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2821 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2881 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2941 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3001 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3061 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3121 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3181 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3241 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3301 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3361 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3421 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3481 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3541 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3601 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3661 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3721 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3781 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3841 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3901 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3961 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4021 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
