LOCUS       sequence-398132-        4527 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 4527)
  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 4527)
  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-SLC7A2_STOP
FEATURES             Location/Qualifiers
     source          1..4527
                     /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(2084..2183)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2200..2219)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2201..2219)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2224..2240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2224..2240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2342..2361)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2353..3162
                     /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(2420..2439)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3309..3897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3798..3817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4051..4068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4227..4254)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4346..4432)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4481..4503
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4495..4512
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4496..4512
                     /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 ccaccatgat cccttgcagg
      121 gccgccctga ccttcgcccg ctgtctgatc cggagaaaga tcgtgaccct ggactccctg
      181 gaggatacaa agctgtgccg gtgtctgtct accatggacc tgatcgccct gggagtggga
      241 agcacactgg gagcaggcgt gtatgtgctg gcaggagagg tggcaaaggc agattctggc
      301 cccagcatcg tggtgagctt tctgatcgcc gccctggcat ccgtgatggc aggactgtgc
      361 tacgcagagt tcggagcaag ggtgcctaag accggaagcg cctacctgta tacctacgtg
      421 acagtgggcg agctgtgggc ctttatcaca ggctggaacc tgatcctgtc ctatgtgatc
      481 ggcaccagct ccgtggcaag ggcatggtct ggcacattcg acgagctgct gagcaagcag
      541 atcggccagt ttctgaggac ctatttccgc atgaattaca caggcctggc cgagtaccct
      601 gatttctttg ccgtgtgcct gatcctgctg ctggcaggac tgctgagctt tggcgtgaag
      661 gagtccgcct gggtgaacaa ggtgttcacc gccgtgaata tcctggtgct gctgtttgtg
      721 atggtggccg gcttcgtgaa gggcaacgtg gccaattgga agatcagcga ggagtttctg
      781 aagaacatct ctgccagcgc cagggagcca ccttccgaga atggcacctc tatctatggc
      841 gccggcggct ttatgccata cggcttcacc ggcacactgg caggagcagc cacatgcttc
      901 tatgcctttg tgggcttcga ctgtatcgcc accacaggcg aggaggtgag aaaccctcag
      961 aaggccatcc caatcggcat cgtgaccagc ctgctggtgt gctttatggc atacttcggc
     1021 gtgtccgccg ccctgacact gatgatgcct tactatctgc tggatgagaa gagcccactg
     1081 cccgtggcct tcgagtatgt gggctgggga ccagcaaagt acgtggtggc agcaggatcc
     1141 ctgtgcgcac tgtccacctc tctgctgggc tctatctttc ctatgccaag agtgatctac
     1201 gccatggccg aggacggcct gctgttcaag tgtctggccc agatcaattc caagaccaag
     1261 acacccatca tcgccaccct gtctagcgga gcagtggccg ccctgatggc cttcctgttt
     1321 gacctgaagg ccctggtgga tatgatgtcc atcggcacac tgatggccta ctctctggtg
     1381 gccgcctgcg tgctgatcct gaggtatcag ccaggcctgt cctacgacca gcccaagtgt
     1441 tctcctgaga aggatggact gggatcctct ccaagggtga ccagcaagtc cgagtctcag
     1501 gtgacaatgc tgcagcggca gggctttagc atgagaaccc tgttctgtcc atccctgctg
     1561 ccaacacagc agagcgcctc cctggtgtct ttcctggtgg gctttctggc cttcctggtg
     1621 ctgggcctga gcgtgctgac cacatacgga gtgcacgcaa tcaccaggct ggaggcatgg
     1681 agcctggccc tgctggccct gtttctggtg ctgttcgtgg ccatcgtgct gacaatctgg
     1741 agacagcctc agaaccagca gaaggtggcc tttatggtgc cctttctgcc tttcctgcca
     1801 gccttctcca tcctggtgaa tatctatctg atggtgcagc tgtctgccga tacctgggtg
     1861 aggtttagca tctggatggc catcggcttt ctgatctatt tctcttacgg aatcaggcac
     1921 agcctggagg gacacctgag agacgagaac aatgaggagg acgcataccc agataacgtg
     1981 cacgcagcag cagaggagaa gtccgccatc caggccaacg atcaccaccc ccgcaatctg
     2041 agctcccctt tcatctttca cgagaagaca tctgagttct aagacccagc tttcttgtac
     2101 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     2161 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     2221 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2281 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2341 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2401 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2461 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2521 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2581 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2641 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2701 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2761 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2821 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2881 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2941 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3001 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3061 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3121 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3181 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     3241 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4021 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4081 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4141 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     4201 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     4261 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     4321 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     4381 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     4441 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     4501 acgacggcca gtcttaagct cgggccc
//
