LOCUS       sequence-352192-        5802 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 5802)
  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 5802)
  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-SLC12A7_STOP
FEATURES             Location/Qualifiers
     source          1..5802
                     /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(3359..3458)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3475..3494)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3476..3494)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3499..3515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3499..3515)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3617..3636)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3628..4437
                     /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(3695..3714)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4584..5172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5073..5092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5326..5343
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5502..5529)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5621..5707)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5756..5778
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5770..5787
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5771..5787
                     /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 ccaccatgcc taccaacttc
      121 acagtggtgc cagtggaggc acacgcagac ggaggaggcg atgagaccgc agagaggaca
      181 gaggcaccag gaaccccaga gggaccagag cccgagcgcc ctagcccagg cgacggcaac
      241 ccacgggaga attccccctt cctgaacaat gtggaggtgg agcaggagtc cttctttgag
      301 ggcaagaata tggccctgtt tgaggaggag atggattcta accctatggt gagctccctg
      361 ctgaataagc tggccaacta taccaatctg agccagggag tggtggagca cgaggaggac
      421 gaggagtccc ggagaaggga ggcaaaggca ccaagaatgg gcacattcat cggcgtgtac
      481 ctgccctgcc tgcagaacat cctgggcgtg atcctgttcc tgaggctgac ctggatcgtg
      541 ggagtggcag gcgtgctgga gtcttttctg atcgtggcca tgtgctgtac ctgtacaatg
      601 ctgacagcca tctctatgag cgccatcgca accaatggag tggtgccagc aggaggcagc
      661 tactatatga tctcccgctc tctgggacct gagtttggag gagcagtggg actgtgcttc
      721 tacctgggca ccacatttgc cggcgccatg tatatcctgg gcaccatcga gatcttcctg
      781 acatatatca gcccaggcgc cgccatcttt caggcagagg cagcaggagg agaggcagca
      841 gccatgctgc acaacatgcg ggtgtacggc acctgcacac tggtgctgat ggccctggtg
      901 gtgttcgtgg gcgtgaagta tgtgaataag ctggccctgg tgtttctggc ctgcgtggtg
      961 ctgtctatcc tggccatcta cgccggcgtg atcaagagcg ccttcgaccc ccctgatatc
     1021 cccgtgtgcc tgctgggcaa cagaacactg tctcgccgga gctttgatgc ttgcgttaaa
     1081 gcctatggca ttcacaacaa ttccgccacc tctgccctgt ggggcctgtt ctgcaatgga
     1141 agccagccat ccgccgcatg tgacgagtac ttcatccaga acaatgtgac agagatccag
     1201 ggaatccctg gagcagcaag cggcgtgttc ctggagaacc tgtggtccac ctacgcacac
     1261 gcaggagcct ttgtggagaa gaagggagtg ccttccgtgc cagtggcaga ggagtctaga
     1321 gcaagcgccc tgccttacgt gctgacagat atcgccgcct ccttcaccct gctggtgggc
     1381 atctattttc catctgtgac cggcatcatg gccggctcca acaggtctgg cgacctgaag
     1441 gatgcccaga agagcatccc caccggcaca atcctggcca tcgtgaccac atctttcatc
     1501 tacctgagct gcatcgtgct gtttggcgcc tgtatcgagg gcgtggtgct gagggacaag
     1561 ttcggcgagg ccctgcaggg caacctggtc atcggaatgc tggcatggcc aagcccttgg
     1621 gtcatcgtga tcggcagctt cttttccaca tgcggagcag gactgcagtc cctgaccgga
     1681 gcacctagac tgctgcaggc aatcgccagg gatggcatcg tgccattcct gcaggtgttt
     1741 ggacacggca aggcaaacgg agagccaacc tgggcactgc tgctgacagt gctgatctgt
     1801 gagaccggca tcctgatcgc ctctctggac agcgtggccc ccatcctgag catgttcttt
     1861 ctgatgtgct acctgttcgt gaacctggca tgtgcagtgc agaccctgct gaggacaccc
     1921 aattggcgcc ctcggttcaa gttttatcac tggaccctgt ccttcctggg catgtctctg
     1981 tgcctggccc tgatgtttat ctgttcttgg tactatgcac tgagcgccat gctgatcgca
     2041 ggatgcatct acaagtatat cgagtacaga ggcgccgaga aggagtgggg cgatggcatc
     2101 aggggcctgt ccctgaatgc agcaagatat gcactgctga gggtggagca cggaccacca
     2161 cacacaaaga actggagacc acaggtgctg gtcatgctga atctggacgc agagcaggca
     2221 gtgaagcacc caaggctgct gtccttcacc tctcagctga aggccggcaa gggcctgaca
     2281 atcgtgggca gcgtgctgga gggcacctat ctggataagc acatggaggc ccagagagcc
     2341 gaggagaaca tcaggagcct gatgtccacc gagaagacaa agggcttttg ccagctggtg
     2401 gtgtctagct ccctgcggga cggcatgagc cacctgatcc agtccgccgg actgggaggc
     2461 ctgaagcaca atacagtgct gatggcctgg cccgcctctt ggaagcagga ggataaccct
     2521 ttcagctgga agaattttgt ggacaccgtg cgcgatacca cagcagcaca ccaggccctg
     2581 ctggtggcca agaacgtgga ctccttccct cagaatcagg agcggtttgg cggcggccac
     2641 atcgacgtgt ggtggatcgt gcacgatggc ggcatgctga tgctgctgcc attcctgctg
     2701 cgccagcaca aagtgtggcg gaagtgtaga atgaggatct ttaccgtggc ccaggtggac
     2761 gataacagca tccagatgaa gaaggatctg cagatgttcc tgtaccacct gagaatctct
     2821 gccgaagtgg aggtggtgga gatggtggag aatgacatca gcgcctttac ctatgagcgc
     2881 acactgatga tggagcagcg gtcccagatg ctgaagcaga tgcagctgtc taagaacgag
     2941 caggagagag aggcccagct gatccacgac aggaataccg cctcccacac agcagcagca
     3001 gcaaggaccc aggcacctcc aacacccgat aaggtgcaga tgacctggac acgggagaag
     3061 ctgatcgccg agaagtaccg ctctcgggac acaagcctgt ccggcttcaa ggacctgttt
     3121 tccatgaagc ccgatcagtc taacgtgaga aggatgcaca ccgccgtgaa gctgaacggc
     3181 gtggtgctga ataagtccca ggacgcccag ctggtgctgc tgaacatgcc tggaccacct
     3241 aagaacaggc agggcgatga gaattatatg gagttcctgg aggtgctgac agagggcctg
     3301 aatcgggtgc tgctggtgag aggaggagga agagaagtga tcaccatcta cagctaagac
     3361 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     3421 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     3481 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     3541 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     3601 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     3661 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     3721 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     3781 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     3841 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     3901 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     3961 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     4021 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     4081 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     4141 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     4201 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     4261 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     4321 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     4381 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     4441 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     4501 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     4561 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4621 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4681 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4741 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4801 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4861 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4921 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4981 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5041 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5101 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5161 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5221 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5281 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5341 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     5401 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     5461 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     5521 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     5581 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     5641 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     5701 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     5761 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
