LOCUS       sequence-261000-        4440 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 4440)
  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 4440)
  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_SLC6A4
FEATURES             Location/Qualifiers
     source          1..4440
                     /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(2565..2664)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2681..2700)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2682..2700)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2705..2721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2705..2721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2823..2842)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2834..3643
                     /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(2901..2920)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3790..4378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4279..4298
                     /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 accatggaga ccacacctct gaattcccag aagcagctgt ctgcctgcga
      721 ggacggcgag gattgtcagg agaacggcgt gctgcagaag gtggtgccaa cccccggcga
      781 caaggtggag tctggccaga tcagcaatgg ctactccgcc gtgccttctc caggagcagg
      841 cgacgatacc aggcactcta tcccagccac cacaaccaca ctggtggcag agctgcacca
      901 gggagagaga gagacatggg gcaagaaggt ggactttctg ctgagcgtga tcggctatgc
      961 cgtggatctg ggcaacgtgt ggcggtttcc ctatatctgc taccagaacg gaggaggagc
     1021 cttcctgctg ccatacacca tcatggccat cttcggcggc atccccctgt tttatatgga
     1081 gctggccctg ggccagtacc accggaatgg ctgcatcagc atctggagaa agatctgtcc
     1141 catctttaag ggcatcggct atgccatctg tatcatcgcc ttctacatcg cctcctacta
     1201 taacacaatc atggcctggg ccctgtacta tctgatcagc tccttcaccg accagctgcc
     1261 ctggacaagc tgcaagaatt cctggaacac cggcaattgt acaaactact tcagcgagga
     1321 taatatcacc tggacactgc actccacctc tcctgccgag gagttctaca caaggcacgt
     1381 gctgcagatc caccgctcca agggactgca ggatctggga ggcatctctt ggcagctggc
     1441 cctgtgcatc atgctgatct tcaccgtgat ctacttcagc atctggaagg gcgtgaagac
     1501 atccggcaag gtggtgtggg tgaccgccac atttccatac atcatcctga gcgtgctgct
     1561 ggtgagggga gccaccctgc caggagcatg gaggggcgtg ctgttctacc tgaagcctaa
     1621 ctggcagaag ctgctggaga caggcgtgtg gatcgacgca gcagcacaga tcttctttag
     1681 cctgggacca ggattcggcg tgctgctggc ctttgcctcc tataataagt tcaacaataa
     1741 ctgctaccag gatgccctgg tgacctccgt ggtgaactgt atgacaagct tcgtgtccgg
     1801 cttcgtgatc tttaccgtgc tgggctatat ggccgagatg cggaatgagg acgtgtctga
     1861 ggtggccaag gatgccggcc ctagcctgct gtttatcacc tacgccgagg ccatcgccaa
     1921 catgccagcc tccacattct ttgccatcat cttctttctg atgctgatca ccctgggcct
     1981 ggactctaca ttcgccggac tggagggcgt gatcaccgcc gtgctggatg agtttcctca
     2041 cgtgtgggcc aagcggagag agaggttcgt gctggccgtg gtcatcacat gcttctttgg
     2101 ctccctggtg accctgacat tcggcggcgc ctatgtggtg aagctgctgg aggagtacgc
     2161 aaccggacca gccgtgctga cagtggccct gatcgaggcc gtggccgtgt cttggtttta
     2221 tggcatcacc cagttctgta gggacgtgaa ggagatgctg ggcttctctc caggctggtt
     2281 ttggaggatc tgctgggtgg caatcagccc actgttcctg ctgtttatca tctgttcttt
     2341 tctgatgagc ccccctcagc tgaggctgtt ccagtacaat tatccctact ggagcatcat
     2401 cctgggctac tgcatcggca cctctagctt tatctgtatc cctacatata tcgcctaccg
     2461 gctgatcatc accccaggca cattcaagga gagaatcatc aagtccatca cccccgagac
     2521 ccctacagag atcccttgtg gcgatatccg cctgaacgcc gtggacccag ctttcttgta
     2581 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2641 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2701 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2761 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2821 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2881 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2941 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3001 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3061 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3121 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3181 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3241 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3301 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3361 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3421 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3481 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3541 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3601 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3661 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3721 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3781 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3841 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3901 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3961 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4021 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4081 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4141 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4201 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4261 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4321 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4381 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
