LOCUS       sequence-263150-        4896 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 4896)
  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 4896)
  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_SLC45A1
FEATURES             Location/Qualifiers
     source          1..4896
                     /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(3021..3120)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3137..3156)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3138..3156)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3161..3177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3161..3177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3279..3298)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3290..4099
                     /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(3357..3376)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4246..4834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4735..4754
                     /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 accatgctgc agcagccagg acctaggcca ggccgccagc agccctccgg
      721 cgacagggat gcatgcaggc tgcaccctca gggcaggccc cctgccctgc caaccatgat
      781 ccctgcagca agctccacac caccaggcga cgcactgttc ccaagcgtgg caccccagga
      841 tttttggcgc tcccaggtga ccggatacag cggatccgtg acaaggcacc tgtcccaccg
      901 cgccaacaac ttcaagcggc accctaagcg gagaaagtgc atcagaccaa gccctccacc
      961 ccctccaaac accccctgtc ctctggagct ggtggacttc ggcgatctgc acccccagcg
     1021 gtccttcaga gagctgctgt ttaatggctg catcctgttc ggcatcgagt tttcttacgc
     1081 catggagacc gcctatgtga cacccgtgct gctgcagatg ggcctgcctg accagctgta
     1141 tagcctggtg tggttcatct ccccaatcct gggctttctg ctgcagccac tgctgggagc
     1201 atggtctgat aggtgtacca gcagattcgg caggcgccgg ccctttatcc tggtgctggc
     1261 aatcggcgcc ctgctgggac tgagcctgct gctgaacgga agggacatcg gaatcgccct
     1321 ggcagatgtg accggcaatc acaagtgggg actgctgctg acagtgtgcg gagtggtgct
     1381 gatggacttc agcgccgact ccgccgataa cccatctcac gcctacatga tggacgtgtg
     1441 cagccccgcc gaccaggata gaggcctgaa tatccacgca ctgctggcag gactgggagg
     1501 aggcttcggc tacgtggtgg gcggcatcca ctgggataag accggatttg gacgggccct
     1561 gggaggacag ctgagagtga tctatctgtt caccgccgtg acactgtccg tgaccacagt
     1621 gctgacactg gtgtctatcc cagagaggcc actgaggcca ccttctgaga agagggccgc
     1681 catgaagtct cctagcctgc cactgccacc aagccctcca gtgctgcctg aggagggacc
     1741 aggcgactct ctgcctagcc acaccgccac aaacttttct agcccaatct cccccccttc
     1801 tcccctgacc cctaagtacg gcagcttcat ctcccgcgat tcctctctga ccggcatcag
     1861 cgagttcgcc agctcctttg gcacagccaa tatcgactcc gtgctgatcg attgctttac
     1921 aggaggacac gactcctacc tggcaatccc aggatctgtg cctaggccac ccatctccgt
     1981 gtctttccca agggcaccag acggctttta tcggcaggat agaggactgc tggagggaag
     2041 ggagggcgcc ctgaccagcg gctgtgacgg cgatatcctg cgcgtgggct ccctggacac
     2101 ctctaagccc aggtctagcg gcatcctgaa gcgcccacag acactggcaa tcccagatgc
     2161 agcaggagga ggaggccctg agaccagcag aaggcgcaac gtgacattct cccagcaggt
     2221 ggccaacatc ctgctgaatg gcgtgaagta cgagtccgag ctgaccggat cctctgagag
     2281 ggcagagcag cctctgtctg tgggcagact gtgcagcacc atctgtaaca tgccaaaggc
     2341 cctgaggaca ctgtgcgtga atcacttcct gggctggctg agctttgagg gcatgctgct
     2401 gttctatacc gactttatgg gcgaggtggt gtttcagggc gatcctaagg ccccacacac
     2461 ctctgaggcc taccagaagt ataacagcgg cgtgacaatg ggctgctggg gcatgtgcat
     2521 ctacgccttc tccgccgcct tttattctgc catcctggag aagctggagg agttcctgtc
     2581 cgtgcggacc ctgtacttca tcgcctatct ggcctttggc ctgggaaccg gactggccac
     2641 actgtccaga aatctgtacg tggtgctgtc tctgtgcatc acatatggca tcctgtttag
     2701 caccctgtgc acactgccct actccctgct gtgcgactac tatcagtcta agaagttcgc
     2761 aggaagctcc gccgacggaa ccaggagagg catgggcgtg gatatctctc tgctgagctg
     2821 ccagtacttt ctggcccaga tcctggtgag cctggtgctg ggaccactga caagcgccgt
     2881 gggatccgcc aacggcgtga tgtatttctc tagcctggtg tcctttctgg gctgtctgta
     2941 ctcctctctg ttcgtgatct atgagatccc tccaagcgac gccgccgatg aggagcacag
     3001 acctctgctg ctgaatgtgg acccagcttt cttgtacaaa gttggcatta taagaaagca
     3061 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3121 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     3181 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3241 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3301 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3361 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     3421 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3481 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3541 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3601 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3661 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3721 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3781 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3841 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3901 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3961 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     4021 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     4081 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     4141 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     4201 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4261 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4321 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4381 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4441 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4501 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4561 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4621 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4681 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4741 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4801 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4861 gttcctggcc ttttgctggc cttttgctca catgtt
//
