LOCUS       sequence-262894-        4680 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 4680)
  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 4680)
  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_SLC44A4
FEATURES             Location/Qualifiers
     source          1..4680
                     /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(2805..2904)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2921..2940)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2922..2940)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2945..2961)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2945..2961)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3063..3082)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3074..3883
                     /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(3141..3160)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4030..4618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4519..4538
                     /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 accatgggcg gcaagcagag ggatgaggac gatgaggcct acggcaagcc
      721 agtgaagtat gacccctctt tcagaggccc tatcaagaac aggagctgca ccgatgtgat
      781 ctgctgcgtg ctgttcctgc tgtttatcct gggctacatc gtggtgggca tcgtggcctg
      841 gctgtacggc gatccacgcc aggtgctgta tccccggaat agcacaggcg cctactgtgg
      901 catgggcgag aacaaggaca agccctacct gctgtatttc aatatctttt cctgcatcct
      961 gagctccaac atcatctctg tggccgagaa tggcctgcag tgtcctaccc cacaggtgtg
     1021 cgtgtctagc tgtcccgagg acccttggac agtgggcaag aacgagtttt ctcagaccgt
     1081 gggcgaggtg ttctacacaa agaacagaaa tttttgcctg ccaggcgtgc cctggaatat
     1141 gaccgtgatc acaagcctgc agcaggagct gtgccctagc ttcctgctgc cttccgcccc
     1201 agccctgggc aggtgttttc catggaccaa cgtgacacca cctgccctgc caggaatcac
     1261 caatgatacc acaatccagc agggcatctc tggcctgatc gacagcctga acgcccgcga
     1321 tatcagcgtg aagatcttcg aggactttgc ccagtcctgg tactggatcc tggtggccct
     1381 gggagtggcc ctggtgctgt ccctgctgtt catcctgctg ctgaggctgg tggcaggacc
     1441 actggtgctg gtgctgatcc tgggcgtgct gggcgtgctg gcatatggca tctactattg
     1501 ctgggaggag taccgcgtgc tgcgggataa gggcgcctcc atctctcagc tgggcttcac
     1561 cacaaatctg tctgcctatc agagcgtgca ggagacatgg ctggccgccc tgatcgtgct
     1621 ggccgtgctg gaggccatcc tgctgctgat gctgatcttt ctgcggcaga gaatcaggat
     1681 cgccatcgcc ctgctgaagg aggccagcaa ggccgtgggc cagatgatgt ccaccatgtt
     1741 ctaccccctg gtgacatttg tgctgctgct gatctgtatc gcctactggg ccatgaccgc
     1801 cctgtatctg gccacatctg gccagcctca gtatgtgctg tgggccagca acatctcctc
     1861 tcctggctgc gagaaggtgc caatcaacac ctcctgtaat cctacagccc acctggtgaa
     1921 tagctcctgc ccaggcctga tgtgcgtgtt ccagggctat tctagcaagg gcctgatcca
     1981 gcggagcgtg ttcaacctgc agatctacgg cgtgctgggc ctgttttgga ccctgaattg
     2041 ggtgctggcc ctgggacagt gcgtgctggc aggagccttc gccagctttt attgggcctt
     2101 ccacaagcca caggatatcc ccaccttccc tctgatctcc gcctttatca gaaccctgag
     2161 gtaccacaca ggctctctgg cctttggcgc cctgatcctg acactggtgc agatcgccag
     2221 agtgatcctg gagtatatcg accacaagct gaggggagtg cagaacccag tggcacggtg
     2281 catcatgtgc tgtttcaagt gctgtctgtg gtgtctggag aagttcatca agtttctgaa
     2341 cagaaatgcc tacatcatga tcgccatcta tggcaagaac ttctgcgtga gcgccaagaa
     2401 tgcctttatg ctgctgatgc gcaacatcgt gcgggtggtg gtgctggaca aggtgaccga
     2461 tctgctgctg ttctttggca agctgctggt ggtgggagga gtgggcgtgc tgtccttctt
     2521 tttcttttct ggcaggatcc ctggcctggg caaggatttc aagagccccc acctgaatta
     2581 ctattggctg cctatcatga cctccatcct gggcgcctac gtgatcgcct ccggcttctt
     2641 cagcgtgttc ggcatgtgcg tggacacact gttcctgtgc ttcctggagg atctggagag
     2701 aaacaatggc agcctggaca ggccatacta tatgagcaag tccctgctga agatcctggg
     2761 caagaagaac gaggccccac ccgacaataa gaagcgcaag aaggacccag ctttcttgta
     2821 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2881 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2941 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     3001 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     3061 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     3121 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3181 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3241 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3301 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3361 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3421 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3481 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3541 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3601 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3661 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3721 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3781 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3841 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3901 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3961 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4021 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4081 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4141 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4201 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4261 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4321 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4381 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4441 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4501 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4561 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4621 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
