LOCUS       sequence-352160-        4608 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 4608)
  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 4608)
  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-SLC3A1_STOP
FEATURES             Location/Qualifiers
     source          1..4608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             312..320
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(2165..2264)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2281..2300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2282..2300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2305..2321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2305..2321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2423..2442)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2434..3243
                     /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(2501..2520)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3390..3978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3879..3898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4132..4149
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4308..4335)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4427..4513)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4562..4584
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4576..4593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4577..4593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc cgaggataag
      121 tctaagaggg acagcatcga gatgtccatg aagggctgcc agaccaacaa tggcttcgtg
      181 cacaacgagg atatcctgga gcagacacca gatcccggca gctccaccga caatctgaag
      241 cacagcacaa ggggaatcct gggatcccag gagccagact tcaagggagt gcagccatac
      301 gcaggaatgc ctaaggaggt gctgttccag ttttccggcc aggcccggta tagaatcccc
      361 agagagatcc tgttttggct gaccgtggca tccgtgctgg tgctgatcgc agcaacaatc
      421 gccatcatcg ccctgtctcc taagtgtctg gactggtggc aggagggccc tatgtaccag
      481 atctatccac gctctttcaa ggacagcaac aaggatggca atggcgacct gaagggcatc
      541 caggacaagc tggattacat caccgccctg aacatcaaga cagtgtggat caccagcttt
      601 tacaagtcta gcctgaagga tttcaggtac ggcgtggagg attttcgcga ggtggaccca
      661 atcttcggca ccatggagga ctttgagaac ctggtggccg ccatccacga taagggcctg
      721 aagctgatca tcgacttcat ccccaatcac acctctgata agcacatctg gtttcagctg
      781 agcaggacac gcaccggcaa gtacacagat tactatatct ggcacgactg cacacacgag
      841 aacggcaaga ccatcccccc taacaattgg ctgtccgtgt acggcaactc ctcttggcac
      901 ttcgacgagg tgcggaatca gtgttatttc caccagttta tgaaggagca gcccgatctg
      961 aacttccgga atcctgacgt gcaggaggag atcaaggaga tcctgagatt ttggctgacc
     1021 aagggcgtgg atggcttctc cctggacgcc gtgaagtttc tgctggaggc caagcacctg
     1081 agagatgaga tccaagtgaa taagacacag atccctgaca cagtgaccca gtactctgag
     1141 ctgtatcacg acttcaccac aacccaagtg ggcatgcacg atatcgtgag gtcctttcgc
     1201 cagacaatgg accagtactc taccgagcca ggccggtata gattcatggg caccgaggcc
     1261 tacgccgagt ctatcgatag gacagtgatg tactatggcc tgcccttcat ccaggaggcc
     1321 gatttcccct ttaacaatta cctgtctatg ctggacaccg tgtctggcaa cagcgtgtat
     1381 gaagtgatca caagctggat ggagaacatg ccagagggca agtggcccaa ttggatgatc
     1441 ggaggaccag acagctccag gctgaccagc agactgggca atcagtacgt gaacgtgatg
     1501 aatatgctgc tgttcacact gcctggcaca ccaatcacct actatggcga ggagatcggc
     1561 atgggcaaca tcgtggccgc caacctgaat gagagctatg atatcaatac cctgaggagc
     1621 aagtccccta tgcagtggga caactctagc aatgccggct tttccgaggc ctctaacaca
     1681 tggctgccta ccaattccga ttaccacacc gtgaacgtgg acgtgcagaa gacacagcca
     1741 cgcagcgccc tgaagctgta tcaggatctg tccctgctgc acgccaacga gctgctgctg
     1801 aatcggggct ggttctgcca cctgagaaac gacagccact acgtggtgta taccagggag
     1861 ctggacggca tcgatcgcat cttcatcgtg gtgctgaatt ttggcgagtc tacactgctg
     1921 aacctgcaca atatgatcag cggactgcca gcaaagatga ggatcaggct gagcaccaac
     1981 tccgccgata agggctccaa ggtggacaca tctggcatct tcctggataa gggcgagggc
     2041 ctgatctttg agcacaacac caagaatctg ctgcacaggc agacagcctt ccgggacaga
     2101 tgctttgtga gcaaccgcgc ctgttactcc tctgtgctga atatcctgta tacctcctgt
     2161 taagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2221 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     2281 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2341 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     2401 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2461 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2521 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2581 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2641 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2701 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2761 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2821 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2881 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2941 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     3001 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     3061 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     3121 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3181 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3241 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3301 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4021 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4081 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4141 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     4201 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     4261 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     4321 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     4381 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     4441 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     4501 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     4561 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccc
//
