LOCUS       sequence-263145-        4731 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 4731)
  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 4731)
  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_SV2C
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..108
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(267..294)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(386..472)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     521..543
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     535..552
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     536..552
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    568..667
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2855..2954)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2971..2990)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2972..2990)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2995..3011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2995..3011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3113..3132)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3124..3933
                     /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(3191..3210)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4080..4668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4569..4588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
       61 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      121 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      181 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      241 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
      301 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
      361 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
      421 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
      481 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
      541 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
      601 tgttcgttgc aacacattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      661 gcaggctcca ccatggagga tagctacaag gacaggacct ccctgatgaa gggcgccaag
      721 gatatcgccc gcgaggtgaa gaagcagacc gtgaagaagg tgaaccaggc cgtggatcgg
      781 gcccaggacg agtatacaca gcggagctac tccagattcc aggacgagga ggacgatgac
      841 gattactatc cagcaggaga gacctataac ggagaggcaa atgacgatga gggaagctcc
      901 gaggcaacag agggacacga tgaggacgat gagatctatg agggcgagta ccagggcatc
      961 ccaagcatga atcaggccaa ggactctatc gtgagcgtgg gccagcccaa gggcgatgag
     1021 tacaaggaca ggagagagct ggagtccgag aggagggcag atgaggagga gctggcacag
     1081 cagtatgagc tgatcatcca ggagtgcggc cacggcagat tccagtgggc cctgttcttt
     1141 gtgctgggca tggccctgat ggccgatggc gtggaggtgt tcgtggtggg ctttgtgctg
     1201 cctagcgccg agaccgacct gtgcatccca aactctggca gcggctggct gggctccatc
     1261 gtgtacctgg gcatgatggt gggagccttc ttttggggag gactggcaga caaagtggga
     1321 aggaagcagt ctctgctgat ctgcatgagc gtgaatggct tctttgcctt cctgtctagc
     1381 tttgtgcagg gctacggctt ctttctgttt tgtaggctgc tgagcggatt cggaatcgga
     1441 ggagccatcc ctaccgtgtt ctcctatttt gcagaggtgc tggcaaggga gaagagagga
     1501 gagcacctgt cttggctgtg catgttttgg atgatcggag gaatctacgc aagcgccatg
     1561 gcatgggcaa tcatccccca ctatggctgg tccttctcta tgggctccgc ctaccagttt
     1621 cactcttggc gggtgttcgt gatcgtgtgc gccctgcctt gcgtgagctc tgtggtggcc
     1681 ctgaccttta tgcccgagag ccctagattc ctgctggaag tgggcaagca cgatgaggcc
     1741 tggatgatcc tgaagctgat ccacgacaca aacatgcggg ccagaggcca gcccgagaag
     1801 gtgtttaccg tgaataagat caagacacct aagcagatcg atgagctgat cgagatcgag
     1861 tccgacaccg gcacatggta tcggagatgc ttcgtgagga tccgcaccga gctgtacggc
     1921 atctggctga cattcatgcg gtgttttaac tatccagtga gagacaatac catcaagctg
     1981 acaatcgtgt ggttcaccct gtcttttggc tactatggcc tgagcgtgtg gttcccagat
     2041 gtgatcaagc ccctgcagag cgacgagtac gccctgctga ccaggaacgt ggagcgcgat
     2101 aagtatgcca acttcaccat caacttcacc atggagaacc agatccacac aggcatggag
     2161 tacgacaatg gcaggttcat cggcgtgaag tttaagtctg tgaccttcaa ggatagcgtg
     2221 ttcaagagct gcacattcga ggacgtgacc tccgtgaaca catactttaa gaattgtacc
     2281 ttcatcgata cagtgtttga taacaccgac ttcgagccat ataagtttat cgacagcgag
     2341 ttcaagaact gctccttctt tcacaataag accggctgtc agatcacatt tgacgatgac
     2401 tacagcgcct attggatcta cttcgtgaac tttctgggca cactggccgt gctgcccggc
     2461 aatatcgtgt ccgccctgct gatggatagg atcggccgcc tgaccatgct gggaggatct
     2521 atggtgctgt ccggcatctc ttgcttcttt ctgtggttcg gcacaagcga gtccatgatg
     2581 atcggcatgc tgtgcctgta caacggcctg accatctccg cctggaattc tctggatgtg
     2641 gtgacagtgg agctgtatcc taccgacagg agggcaacag gattcggctt tctgaacgca
     2701 ctgtgcaagg cagcagccgt gctgggcaat ctgatcttcg gcagcctggt gtccatcacc
     2761 aagtccatcc ctatcctgct ggcctctaca gtgctggtgt gcggaggact ggtgggactg
     2821 tgcctgccag acaccagaac acaggtgctg atggacccag ctttcttgta caaagttggc
     2881 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     2941 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     3001 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     3061 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     3121 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     3181 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     3241 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     3301 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     3361 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     3421 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     3481 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     3541 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     3601 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     3661 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     3721 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     3781 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3841 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3901 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     3961 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     4021 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt c
//
