LOCUS       sequence-442633-        5370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for gateway cloning of HyPer7 DAAO, a fusion protein 
            used to measure the uptake of D-amino acids across the plasma 
            membrane..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5370)
  AUTHORS   Johal AS, Al-Shekaili HH, Abedrabbo M, Kehinde AZ, Towriss M, Koe 
            JC, Hewton KG, Thomson SB, Ciernia AV, Leavitt B, Parker SJ
  TITLE     Restricting lysine normalizes toxic catabolites associated with 
            ALDH7A1 deficiency in cells and mice.
  JOURNAL   Cell Rep. 2024 Dec 24;43(12):115069. doi: 
            10.1016/j.celrep.2024.115069. Epub 2024 Dec 10.
  PUBMED    39661514
REFERENCE   2  (bases 1 to 5370)
  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"; journalName: "Cell Rep.";
            date: "2024-12-24"; volume: "43(12):115069. doi"; pages: " 
            10.1016/j.celrep.2024.115069. Epub 2024 Dec 10"
COMMENT     Sequence Label: pDONR223_HyPer7RgDAAO
FEATURES             Location/Qualifiers
     source          1..5370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     1285..1306
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1415..1436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1676..1695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             3218..3244
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     protein_bind    complement(3249..3348)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3365..3384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3366..3384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3389..3405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3389..3405)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3836..4627
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4720..5308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5209..5228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc accatgcacc tggctaatga ggagcaaaac gaattggaag gtgcttttaa
      721 gctgggcttg atctttaccg tcgccccgta cttgctgcct aagttgatcg tcagcctgcg
      781 ccgcaccgcc ccgaagatgc ctttgatgtt ggaggagaat tacacccaca cccttaccga
      841 gtcactgaaa cgcggcgacg ttgacgccat cattgtggca gagccgtttc aagagccggg
      901 tattgtgact gaacctttgt atgacgagcc gtttttcgtt atcgttccga agggccatag
      961 cttcgaggag ttggatgccg ttagccctcg catgttgggc gaagaacaag ttctgcttct
     1021 gaccgagggc aactgtatgc gtgaccaggt gcttagctcc gccggcttca acagcaacaa
     1081 cgtctatatc atggccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     1141 caacgtcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1201 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc ttccagtccg tcctgagcaa
     1261 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1321 cactctcggc atggacgagc tgtacaacgt ggatggcggt agcggtggca ccggcagcaa
     1381 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1441 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1501 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1561 cctcagctac ggcctgaagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1621 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1681 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1741 cgagctgaag ggcatcggct tcaaggagga cggcaacatc ctggggcaca agctggagta
     1801 caacggcccc agttgcagcg agcttgccgc caaacagcgc attcagggtc tcaccaacac
     1861 ccttcaaggc tcaagcatca ataccatccg ccacatggtt gccagtggct tggcaatcag
     1921 cgtcttgcct gccactgccc ttaccgaaaa cgaccacatg ctgtttagca ttatcccgtt
     1981 tgaaggcacc ccgcctagcc gccgcgttgt cctggcctat cgccgcaact ttgtccgccc
     2041 taaggccctt tcagccatga aagccgcaat tatgcaaagc caactgcatg gtgttagctt
     2101 catctgcgat ggtggtagtg gtcacagcca gaagagggtg gtcgtgctgg gctccggagt
     2161 catcgggctc tctagcgccc tgattctcgc tagaaaaggc tactccgtgc atatcctggc
     2221 cagagacttg cctgaggatg tcagctccca gaccttcgca agcccctggg ccggagctaa
     2281 ctggacacca tttatgaccc tgacagacgg acccagacaa gccaagtggg aagagtccac
     2341 cttcaagaaa tgggtggagc tggttcccac aggccacgct atgtggctga aagggaccag
     2401 aaggtttgcc cagaatgagg atggcctcct gggacattgg tacaaggaca tcacccctaa
     2461 ctataggcca cttccaagct ctgaatgtcc acctggagcc attggagtga catacgacac
     2521 cctgagcgtc cacgctccca agtattgcca ataccttgcc agagagctgc agaaacttgg
     2581 ggctaccttc gaaaggagaa cagtgactag cctggagcaa gccttcgatg gagctgacct
     2641 cgtggtcaac gccaccgggc tgggagctaa gtccattgcc ggcatcgatg accaggctgc
     2701 cgagcccatt aggggacaga ccgtgcttgt caaatctccc tgtaagaggt gcacaatgga
     2761 tagctccgac cctgcttctc ccgcttatat cattcctaga ccaggaggag aagtgatctg
     2821 tggcggaacc tacggagtcg gcgattggga cctgagcgtg aatcccgaaa cagtccaaag
     2881 gattctcaag cactgcctga gacttgaccc taccattagc tccgatggaa ctatcgaggg
     2941 cattgaagtg ctgagacata acgttggcct gagacccgcc aggagaggag gacctagggt
     3001 ggaggctgaa agaatcgtct tgcccctgga ccggaccaag agccctctct ctctgggaag
     3061 agggtccgca cgggctgcaa aagagaagga agtgaccctt gtccacgcct acggctttag
     3121 tagcgcaggg tatcagcaat cctggggagc cgctgaggac gtggctcagc tggtcgatga
     3181 agccttccaa agataccacg gcgcagctag ggagagcctg ccccccctgg agcggctgac
     3241 cctgtaaaac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     3301 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     3361 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3421 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     3481 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     3541 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     3601 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     3661 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     3721 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     3781 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     3841 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     3901 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     3961 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     4021 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     4081 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     4141 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     4201 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     4261 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     4321 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     4381 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     4441 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     4501 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     4561 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     4621 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4861 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5101 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5341 ggccttttgc tggccttttg ctcacatgtt
//
