LOCUS       sequence-345399-        4497 bp    DNA     circular SYN 15-APR-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 4497)
  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 4497)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SLC2A13_STOP
FEATURES             Location/Qualifiers
     source          1..4497
                     /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(2054..2153)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2170..2189)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2171..2189)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2194..2210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2194..2210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2312..2331)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2323..3132
                     /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(2390..2409)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3279..3867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3768..3787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4021..4038
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4197..4224)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4316..4402)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4451..4473
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4465..4482
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4466..4482
                     /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 ccaccatgtc tagaaaggcc
      121 agcgagaacg tggagtacac cctgaggagc ctgagctccc tgatgggcga gcggagaagg
      181 aagcagcctg agccagacgc agcaagcgcc gcaggcgagt gttccctgct ggcagcagca
      241 gagtctagca ccagcctgca gtccgccgga gcaggaggag gaggagtggg cgacctggag
      301 cgcgccgccc gccggcagtt ccagcaggat gagacccctg ccttcgtgta cgtggtggcc
      361 gtgttttccg ccctgggagg cttcctgttt ggctatgaca caggcgtggt gtctggcgcc
      421 atgctgctgc tgaagcggca gctgagcctg gatgccctgt ggcaggagct gctggtgtcc
      481 tctaccgtgg gagcagcagc cgtgtctgcc ctggcaggcg gcgccctgaa tggcgtgttc
      541 ggcagaaggg cagcaatcct gctggcatct gccctgttta ccgcaggaag cgccgtgctg
      601 gcagcagcaa acaataagga gacactgctg gcaggcagac tggtggtggg actgggaatc
      661 ggaatcgcat ccatgaccgt gcccgtgtac atcgccgagg tgtctccccc taacctgaga
      721 ggcaggctgg tgaccatcaa tacactgttc atcacaggcg gccagttctt tgcctccgtg
      781 gtggacggcg ccttttctta cctgcagaag gatggctgga ggtatatgct gggactggca
      841 gcagtgcctg ccgtgatcca gttctttggc ttcctgtttc tgcctgagag cccaagatgg
      901 ctgatccaga agggacagac ccagaaggca aggaggatcc tgagccagat gaggggcaac
      961 cagacaatcg acgaggagta cgattccatc aagaacaata tcgaggagga ggagaaggaa
     1021 gtgggatccg ccggaccagt gatctgcaga atgctgtctt atccacccac cagaagggcc
     1081 ctgatcgtgg gctgtggcct gcagatgttc cagcagctgt ccggcatcaa caccatcatg
     1141 tactatagcg ccacaatcct gcagatgtcc ggcgtggagg acgataggct ggccatctgg
     1201 ctggcctctg tgaccgcctt cacaaatttc atctttaccc tggtgggcgt gtggctggtg
     1261 gagaaagtgg gccggagaaa gctgacattt ggcagcctgg caggaaccac agtggccctg
     1321 atcatcctgg ccctgggatt cgtgctgtcc gcccaggtgt ctccccgcat cacctttaag
     1381 ccaatcgcac ctagcggaca gaacgcaacc tgcacacggt actcctattg caatgagtgt
     1441 atgctggacc ctgattgcgg cttctgttac aagatgaaca agtctaccgt gatcgacagc
     1501 tcctgcgtgc cagtgaacaa ggcaagcaca aatgaggcag catggggccg ctgtgagaac
     1561 gagaccaagt ttaagacaga ggatatcttc tgggcctaca atttttgccc caccccttat
     1621 tcctggacag ccctgctggg cctgatcctg tacctggtgt tctttgcacc aggaatggga
     1681 ccaatgccat ggaccgtgaa ctctgagatc tatccactgt gggcacggag cacaggaaat
     1741 gcatgttcta gcggcatcaa ctggatcttc aatgtgctgg tgagcctgac ctttctgcac
     1801 acagccgagt acctgaccta ctatggcgcc ttctttctgt atgcaggatt cgcagcagtg
     1861 ggactgctgt ttatctacgg ctgcctgccc gagacaaagg gcaagaagct ggaggagatc
     1921 gagtccctgt tcgacaatag actgtgcacc tgtggcacat ccgactctga tgagggccgc
     1981 tacatcgagt atatccgggt gaagggcagc aactatcacc tgtccgacaa tgatgcctct
     2041 gatgtggagt aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     2101 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     2161 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     2221 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2281 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     2341 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2401 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2461 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2521 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2581 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2641 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2701 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2761 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2821 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     2881 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2941 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3001 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3061 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3121 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     3181 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     3241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3421 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3901 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3961 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4021 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4081 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4141 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     4201 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     4261 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     4321 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     4381 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     4441 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
