LOCUS       sequence-362127-        4443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  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 4443)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2000..2099)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2116..2135)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2117..2135)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2140..2156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2140..2156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2258..2277)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2269..3078
                     /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(2336..2355)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3225..3813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3714..3733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3967..3984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4143..4170)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4262..4348)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4397..4419
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4411..4428
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4412..4428
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgga gaccacacct
      121 ctgaattccc agaagcagct gtctgcctgc gaggacggcg aggattgtca ggagaacggc
      181 gtgctgcaga aggtggtgcc aacccccggc gacaaggtgg agtctggcca gatcagcaat
      241 ggctactccg ccgtgccttc tccaggagca ggcgacgata ccaggcactc tatcccagcc
      301 accacaacca cactggtggc agagctgcac cagggagaga gagagacatg gggcaagaag
      361 gtggactttc tgctgagcgt gatcggctat gccgtggatc tgggcaacgt gtggcggttt
      421 ccctatatct gctaccagaa cggaggagga gccttcctgc tgccatacac catcatggcc
      481 atcttcggcg gcatccccct gttttatatg gagctggccc tgggccagta ccaccggaat
      541 ggctgcatca gcatctggag aaagatctgt cccatcttta agggcatcgg ctatgccatc
      601 tgtatcatcg ccttctacat cgcctcctac tataacacaa tcatggcctg ggccctgtac
      661 tatctgatca gctccttcac cgaccagctg ccctggacaa gctgcaagaa ttcctggaac
      721 accggcaatt gtacaaacta cttcagcgag gataatatca cctggacact gcactccacc
      781 tctcctgccg aggagttcta cacaaggcac gtgctgcaga tccaccgctc caagggactg
      841 caggatctgg gaggcatctc ttggcagctg gccctgtgca tcatgctgat cttcaccgtg
      901 atctacttca gcatctggaa gggcgtgaag acatccggca aggtggtgtg ggtgaccgcc
      961 acatttccat acatcatcct gagcgtgctg ctggtgaggg gagccaccct gccaggagca
     1021 tggaggggcg tgctgttcta cctgaagcct aactggcaga agctgctgga gacaggcgtg
     1081 tggatcgacg cagcagcaca gatcttcttt agcctgggac caggattcgg cgtgctgctg
     1141 gcctttgcct cctataataa gttcaacaat aactgctacc aggatgccct ggtgacctcc
     1201 gtggtgaact gtatgacaag cttcgtgtcc ggcttcgtga tctttaccgt gctgggctat
     1261 atggccgaga tgcggaatga ggacgtgtct gaggtggcca aggatgccgg ccctagcctg
     1321 ctgtttatca cctacgccga ggccatcgcc aacatgccag cctccacatt ctttgccatc
     1381 atcttctttc tgatgctgat caccctgggc ctggactcta cattcgccgg actggagggc
     1441 gtgatcaccg ccgtgctgga tgagtttcct cacgtgtggg ccaagcggag agagaggttc
     1501 gtgctggccg tggtcatcac atgcttcttt ggctccctgg tgaccctgac attcggcggc
     1561 gcctatgtgg tgaagctgct ggaggagtac gcaaccggac cagccgtgct gacagtggcc
     1621 ctgatcgagg ccgtggccgt gtcttggttt tatggcatca cccagttctg tagggacgtg
     1681 aaggagatgc tgggcttctc tccaggctgg ttttggagga tctgctgggt ggcaatcagc
     1741 ccactgttcc tgctgtttat catctgttct tttctgatga gcccccctca gctgaggctg
     1801 ttccagtaca attatcccta ctggagcatc atcctgggct actgcatcgg cacctctagc
     1861 tttatctgta tccctacata tatcgcctac cggctgatca tcaccccagg cacattcaag
     1921 gagagaatca tcaagtccat cacccccgag acccctacag agatcccttg tggcgatatc
     1981 cgcctgaacg ccgtgtaaga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     2041 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     2101 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     2161 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     2221 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     2281 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     2341 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     2401 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     2461 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     2521 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     2581 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     2641 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     2701 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     2761 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     2821 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2881 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2941 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     3001 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     3061 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     3121 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     3181 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3241 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3301 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3361 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3421 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3481 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3541 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3601 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3661 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3721 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3781 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3841 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3901 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3961 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4021 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4081 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     4141 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     4201 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     4261 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     4321 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     4381 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     4441 ccc
//
