LOCUS       sequence-261801-        4407 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 4407)
  AUTHORS   Rebsamen M, Girardi E, Sedlyarov V, Scorzoni S, Papakostas K, 
            Vollert M, Konecka J, Guertl B, Klavins K, Wiedmer T, Superti-Furga 
            G
  TITLE     Gain-of-function genetic screens in human cells identify SLC 
            transporters overcoming environmental nutrient restrictions.
  JOURNAL   Life Sci Alliance. 2022 Sep 16;5(11):e202201404. doi: 
            10.26508/lsa.202201404. Print 2022 Nov.
  PUBMED    36114003
REFERENCE   2  (bases 1 to 4407)
  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"; journalName: "Life Sci 
            Alliance."; date: "2022-09-16"; volume: "5(11):e202201404. doi"; 
            pages: " 10.26508/lsa.202201404. Print 2022 Nov"
COMMENT     Sequence Label: pDONR221_SLC7A3
FEATURES             Location/Qualifiers
     source          1..4407
                     /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(2532..2631)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2648..2667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2649..2667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2672..2688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2672..2688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2790..2809)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2801..3610
                     /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(2868..2887)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3757..4345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4246..4265
                     /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 accatgcctt ggcaggcctt ccggagattt ggccagaagc tggtgaggcg
      721 ccggacactg gagagcggaa tggcagagac cagactggcc aggtgcctgt ccacactgga
      781 tctggtggcc ctgggagtgg gatctaccct gggagcaggc gtgtatgtgc tggcaggaga
      841 ggtggcaaag gacaaggcag gcccctccat cgtgatctgc tttctggtgg ccgccctgag
      901 ctccgtgctg gcaggactgt gctacgcaga gttcggagca agggtgcctc ggtccggatc
      961 tgcctacctg tattcttacg tgaccgtggg cgagctgtgg gcctttacca caggctggaa
     1021 cctgatcctg tcttatgtga tcggaacagc aagcgtggca agggcatggt ctagcgcctt
     1081 cgataacctg atcggcaatc acatcagcaa gaccctgcag ggatccatcg ccctgcacgt
     1141 gccacacgtg ctggcagagt accctgactt ctttgcactg ggactggtgc tgctgctgac
     1201 aggactgctg gccctgggag caagcgagtc cgccctggtg acaaaggtgt ttaccggcgt
     1261 gaatctgctg gtgctgggct ttgtgatgat ctccggcttc gtgaagggcg acgtgcacaa
     1321 ctggaagctg accgaggagg attatgagct ggccatggcc gagctgaatg acacatactc
     1381 tctgggacca ctgggaagcg gaggattcgt gcccttcggc tttgagggaa tcctgagggg
     1441 agcagcaacc tgcttctatg cctttgtggg cttcgattgt atcgcaacca caggagagga
     1501 ggcacagaac ccacagcgga gcatccctat gggcatcgtg atctctctga gcgtgtgctt
     1561 tctggcctac ttcgccgtgt cctctgccct gaccctgatg atgccttact atcagctgca
     1621 gccagagtcc ccactgcctg aggcctttct gtatatcgga tgggcacctg caagatacgt
     1681 ggtggccgtg ggcagcctgt gcgccctgtc cacatctctg ctgggctcca tgtttccaat
     1741 gcccagagtg atctacgcca tggccgagga tggactgctg ttccgcgtgc tggcaaggat
     1801 ccacaccgga acaaggaccc caatcatcgc caccgtggtg tctggcatca tcgccgcctt
     1861 catggccttc ctgtttaagc tgaccgacct ggtggatctg atgagcatcg gcacactgct
     1921 ggcctatagc ctggtgtcca tctgcgtgct gatcctgcgg taccagccag accaggagac
     1981 aaagaccgga gaggaggtgg agctgcagga ggaggccatc accacagaga gcgagaagct
     2041 gaccctgtgg ggcctgttct ttcccctgaa ttccatccct acaccactgt ctggccagat
     2101 cgtgtacgtg tgcagctccc tgctggccgt gctgctgacc gccctgtgcc tggtgctggc
     2161 acagtggtcc gtgccactgc tgtctggcga cctgctgtgg acagccgtgg tggtgctgct
     2221 gctgctgctg atcatcggca tcatcgtggt catctggaga cagccacagt ctagcacccc
     2281 cctgcacttt aaggtgcccg ccctgcctct gctgccactg atgagcatct tcgtgaacat
     2341 ctatctgatg atgcagatga cagcaggaac ctgggcaagg ttcggcgtgt ggatgctgat
     2401 cggctttgcc atctatttcg gctacggcat ccagcacagc ctggaggaga tcaagtccaa
     2461 tcagccttct agaaagagca gggccaagac agtggacctg gatccaggca ccctgtacgt
     2521 gcactccgtg gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2581 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2641 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2701 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2761 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2821 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2881 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2941 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3001 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3061 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3121 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3181 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3241 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3301 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3361 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3421 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3481 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3541 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3601 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3661 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3721 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3781 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3841 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3901 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3961 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4021 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4081 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4141 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4201 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4261 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4321 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4381 cttttgctgg ccttttgctc acatgtt
//
