LOCUS       sequence-259281-        4521 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 4521)
  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 4521)
  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_SLC44A1
FEATURES             Location/Qualifiers
     source          1..4521
                     /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(2646..2745)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2762..2781)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2763..2781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2786..2802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2786..2802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2904..2923)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2915..3724
                     /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(2982..3001)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3871..4459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4360..4379
                     /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 accatgggat gctgtagctc cgcctctagc gccgcccagt cctctaagag
      721 ggagtggaag cctctggagg accgctcctg caccgatatc ccatggctgc tgctgttcat
      781 cctgttttgc atcggcatgg gcttcatctg tggcttttct atcgcaacag gagcagcagc
      841 aaggctggtg agcggctacg actcctatgg caacatctgc ggccagaaga ataccaagct
      901 ggaggccatc ccaaacagcg gcatggatca cacacagaga aagtacgtgt tctttctgga
      961 cccctgtaac ctggatctga tcaatcggaa gatcaagagc gtggccctgt gcgtggcagc
     1021 atgtcccaga caggagctga agaccctgtc cgatgtgcag aagttcgccg agatcaacgg
     1081 ctctgccctg tgcagctaca atctgaagcc tagcgagtat accacatccc caaagagcag
     1141 cgtgctgtgc ccaaagctgc cagtgcctgc atccgcccca atccccttct ttcacaggtg
     1201 cgcccctgtg aacatctctt gttacgccaa gttcgccgag gccctgatca ccttcgtgag
     1261 cgacaattct gtgctgcacc gcctgatctc tggcgtgatg acaagcaagg agatcatcct
     1321 gggcctgtgc ctgctgtccc tggtgctgtc tatgatcctg atggtcatca tccggtacat
     1381 cagcagagtg ctggtgtgga tcctgaccat cctggtcatc ctgggatccc tgggaggaac
     1441 aggcgtgctg tggtggctgt atgccaagca gcggagaagc cccaaggaga ccgtgacacc
     1501 tgagcagctg cagatcgccg aggataacct gcgggccctg ctgatctacg ccatctccgc
     1561 caccgtgttc acagtgatcc tgtttctgat catgctggtc atgaggaagc gcgtggccct
     1621 gaccatcgcc ctgttccacg tggccggcaa ggtgtttatc cacctgccac tgctggtgtt
     1681 ccagcccttt tggaccttct ttgccctggt gctgttctgg gtgtattgga tcatgacact
     1741 gctgtttctg ggcaccacag gctctcccgt gcagaatgag cagggcttcg tggagtttaa
     1801 gatcagcggc cctctgcagt acatgtggtg gtatcacgtg gtgggcctga tctggatctc
     1861 cgagttcatc ctggcatgtc agcagatgac cgtggcagga gcagtggtga cctactattt
     1921 cacacgggac aagagaaacc tgccttttac accaatcctg gcctctgtga ataggctgat
     1981 caggtaccac ctgggaaccg tggcaaaggg cagctttatc atcacactgg tgaagatccc
     2041 acggatgatc ctgatgtata tccactccca gctgaagggc aaggagaacg cctgcgccag
     2101 atgcgtgctg aagtcttgca tctgctgtct gtggtgcctg gagaagtgtc tgaattacct
     2161 gaaccagaat gcctataccg ccacagccat caactccacc aatttctgta catctgccaa
     2221 ggacgccttt gtgatcctgg tggagaacgc cctgagggtg gccaccatca atacagtggg
     2281 cgatttcatg ctgtttctgg gcaaggtgct gatcgtgtgc agcaccggac tggcaggaat
     2341 catgctgctg aattaccagc aggattatac agtgtgggtg ctgcccctga tcatcgtgtg
     2401 cctgttcgcc tttctggtgg cccactgttt cctgtccatc tacgagatgg tggtggacgt
     2461 gctgttcctg tgctttgcca tcgacaccaa gtacaacgat ggctctcctg gccgcgagtt
     2521 ctatatggat aaggtgctga tggagtttgt ggagaatagc aggaaggcca tgaaggaggc
     2581 aggcaaggga ggagtggcag acagcaggga gctgaagcca atggcatccg gagcctctag
     2641 cgccgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2701 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2761 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2821 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2881 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2941 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     3001 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     3061 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     3121 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3181 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3241 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3301 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3361 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3421 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3481 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3541 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3601 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3661 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3721 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3781 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3841 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3901 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3961 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4021 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4081 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4141 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4201 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4261 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4321 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4381 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4441 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4501 ctggcctttt gctcacatgt t
//
