LOCUS       sequence-261090-        4854 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 4854)
  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 4854)
  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_SLC45A4
FEATURES             Location/Qualifiers
     source          1..4854
                     /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(2979..3078)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3095..3114)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3096..3114)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3119..3135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3119..3135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3237..3256)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3248..4057
                     /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(3315..3334)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4204..4792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4693..4712
                     /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 accatgaaga tggctcccca gaacgctgat cccgagtcta tgcaggtcca
      721 ggaactgtcc gtcccactgc ccgaccccca gaaagcagga ggcgccgagg ccgagaactg
      781 cgagaccatc tctgagggct ccatcgacag aatccccatg agactgtggg taatgcacgg
      841 cgccgtgatg ttcggccgcg agttttgtta cgccatggag accgccctgg tgacccctat
      901 cctgctgcag atcggcctgc cagagcagta ttactccctg acctggttcc tgtccccaat
      961 cctgggcctg atcttcacac ccctgatcgg cagcgcctct gataggtgca cactgtcttg
     1021 gggcagacgg agacctttca tcctggccct gtgcgtgggc gtgctgtttg gcgtggccct
     1081 gtttctgaat ggctctgcca tcggcctggc cctgggcgac gtgcccaaca gacagcctat
     1141 cggcatcgtg ctgacagtgc tgggcgtggt ggtgctggac ttttccgccg atgccacaga
     1201 gggcccaatc agagcctacc tgctggacgt ggtggattcc gaggagcagg acatggccct
     1261 gaatatccac gcctttagcg ccggcctggg cggcgccatc ggctatgtgc tgggcggcct
     1321 ggattggacc cagacatttc tgggctcttg gtttagaacc cagaatcagg tgctgttctt
     1381 cttcgccgcc atcatcttca cagtgtccgt ggccctgcac ctgttttcca tcgatgagga
     1441 gcagtattcc cctcagcagg agagatctgc cgaggagccc ggcgccctgg atggcggcga
     1501 gccccacggc gtgcctgcct ttcctgacga ggtgcagagc gagcacgagc tggccctgga
     1561 ctatccagat gtggacatca tgaggtccaa gtccgatagc gccctgcacg tgcccgatac
     1621 agccctggat ctggagccag agctgctgtt cctgcacgat atcgagccat ctatcttcca
     1681 cgatgcctcc taccccgcca ccccaagaag cacctctcag gagctggcca agaccaagct
     1741 gccaaggctg gccacctttc tgaaggaggc cgccaaggag gatgagacac tgctggacaa
     1801 ccacctgaac gaggccaagg tgccaaacgg cagcggctct ccaaccaagg atgccctggg
     1861 cggctatacc cgggtggata ccaagccatc tgccacatct agctccatga gacggagaag
     1921 acacgccttc agacggcagg cctcctctac attcagctat tacggcaagc tgggctccca
     1981 ctgctataga taccggagag ccaatgccgt ggtgctgatc aagccttcca ggagcatgtc
     2041 cgacctgtat gacatgcaga agagacagag acagcacagg cacagaaacc agtctggcgc
     2101 caccacatcc agcggcgaca ccgagagcga ggagggcgag ggcgagacaa ccgtgagact
     2161 gctgtggctg tctatgctga agatgccaag agagctgatg aggctgtgcc tgtgccacct
     2221 gctgacctgg tttagcgtga tcgccgaggc cgtgttttat accgacttta tgggccaggt
     2281 aatcttcgag ggcgatccta aggccccaag caattccacc gcctggcagg cctataacgc
     2341 cggcgtgaag atgggctgct ggggcctggt aatctatgcc gccacaggcg ccatctgttc
     2401 cgccctgctg cagaagtacc tggacaatta tgatctgagc gtgagagtga tctacgtgct
     2461 gggcacactg ggctttagcg tgggcaccgc cgtgatggcc atgttcccca acgtgtacgt
     2521 ggccatggtg accatcagca caatgggcat cgtgagcatg tccatcagct actgcccata
     2581 tgccctgctg ggccagtatc acgacatcaa gcagtatatc caccactctc ccggcaatag
     2641 caagcggggc tttggcatcg attgcgccat cctgtcctgt caggtgtaca tctcccagat
     2701 cctggtggcc tccgccctgg gcggcgtggt ggatgccgtg ggcaccgtga gagtgatccc
     2761 tatggtggcc tccgtgggct cttttctggg cttcctgacc gccacattcc tggtaatcta
     2821 ccctaatgtg agcgaggagg ccaaggagga gcagaagggc ctgtcttccc ctctggccgg
     2881 cgagggcagg gccggcggca actctgagaa gcccacagtg ctgaaactga caagaaagga
     2941 agggctgcag ggacccgtgg aaacagaatc cgtcgtcgac ccagctttct tgtacaaagt
     3001 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     3061 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     3121 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     3181 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     3241 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     3301 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     3361 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     3421 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     3481 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     3541 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     3601 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     3661 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     3721 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3781 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3841 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3901 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3961 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     4021 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     4081 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     4141 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4201 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4261 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4321 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4381 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4441 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4501 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4561 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4621 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4681 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4741 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4801 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
