LOCUS       sequence-261195-        4605 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 4605)
  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 4605)
  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
FEATURES             Location/Qualifiers
     source          1..4605
                     /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"
     RBS             880..888
                     /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(2730..2829)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2846..2865)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2847..2865)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2870..2886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2870..2886)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2988..3007)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2999..3808
                     /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(3066..3085)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3955..4543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4444..4463
                     /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 aggataagtc taagagggac agcatcgaga tgtccatgaa
      721 gggctgccag accaacaatg gcttcgtgca caacgaggat atcctggagc agacaccaga
      781 tcccggcagc tccaccgaca atctgaagca cagcacaagg ggaatcctgg gatcccagga
      841 gccagacttc aagggagtgc agccatacgc aggaatgcct aaggaggtgc tgttccagtt
      901 ttccggccag gcccggtata gaatccccag agagatcctg ttttggctga ccgtggcatc
      961 cgtgctggtg ctgatcgcag caacaatcgc catcatcgcc ctgtctccta agtgtctgga
     1021 ctggtggcag gagggcccta tgtaccagat ctatccacgc tctttcaagg acagcaacaa
     1081 ggatggcaat ggcgacctga agggcatcca ggacaagctg gattacatca ccgccctgaa
     1141 catcaagaca gtgtggatca ccagctttta caagtctagc ctgaaggatt tcaggtacgg
     1201 cgtggaggat tttcgcgagg tggacccaat cttcggcacc atggaggact ttgagaacct
     1261 ggtggccgcc atccacgata agggcctgaa gctgatcatc gacttcatcc ccaatcacac
     1321 ctctgataag cacatctggt ttcagctgag caggacacgc accggcaagt acacagatta
     1381 ctatatctgg cacgactgca cacacgagaa cggcaagacc atccccccta acaattggct
     1441 gtccgtgtac ggcaactcct cttggcactt cgacgaggtg cggaatcagt gttatttcca
     1501 ccagtttatg aaggagcagc ccgatctgaa cttccggaat cctgacgtgc aggaggagat
     1561 caaggagatc ctgagatttt ggctgaccaa gggcgtggat ggcttctccc tggacgccgt
     1621 gaagtttctg ctggaggcca agcacctgag agatgagatc caagtgaata agacacagat
     1681 ccctgacaca gtgacccagt actctgagct gtatcacgac ttcaccacaa cccaagtggg
     1741 catgcacgat atcgtgaggt cctttcgcca gacaatggac cagtactcta ccgagccagg
     1801 ccggtataga ttcatgggca ccgaggccta cgccgagtct atcgatagga cagtgatgta
     1861 ctatggcctg cccttcatcc aggaggccga tttccccttt aacaattacc tgtctatgct
     1921 ggacaccgtg tctggcaaca gcgtgtatga agtgatcaca agctggatgg agaacatgcc
     1981 agagggcaag tggcccaatt ggatgatcgg aggaccagac agctccaggc tgaccagcag
     2041 actgggcaat cagtacgtga acgtgatgaa tatgctgctg ttcacactgc ctggcacacc
     2101 aatcacctac tatggcgagg agatcggcat gggcaacatc gtggccgcca acctgaatga
     2161 gagctatgat atcaataccc tgaggagcaa gtcccctatg cagtgggaca actctagcaa
     2221 tgccggcttt tccgaggcct ctaacacatg gctgcctacc aattccgatt accacaccgt
     2281 gaacgtggac gtgcagaaga cacagccacg cagcgccctg aagctgtatc aggatctgtc
     2341 cctgctgcac gccaacgagc tgctgctgaa tcggggctgg ttctgccacc tgagaaacga
     2401 cagccactac gtggtgtata ccagggagct ggacggcatc gatcgcatct tcatcgtggt
     2461 gctgaatttt ggcgagtcta cactgctgaa cctgcacaat atgatcagcg gactgccagc
     2521 aaagatgagg atcaggctga gcaccaactc cgccgataag ggctccaagg tggacacatc
     2581 tggcatcttc ctggataagg gcgagggcct gatctttgag cacaacacca agaatctgct
     2641 gcacaggcag acagccttcc gggacagatg ctttgtgagc aaccgcgcct gttactcctc
     2701 tgtgctgaat atcctgtata cctcctgtga cccagctttc ttgtacaaag ttggcattat
     2761 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2821 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2881 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2941 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     3001 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     3061 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     3121 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     3181 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     3241 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     3301 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3361 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3421 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3481 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3541 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3601 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3661 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3721 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3781 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3841 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3901 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4081 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4561 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
