LOCUS       sequence-338864-        4717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EarlyBac insect cell expression system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4717)
  AUTHORS   Chou TH, Tajima N, Romero-Hernandez A, Furukawa H
  TITLE     Structural Basis of Functional Transitions in Mammalian NMDA 
            Receptors.
  JOURNAL   Cell. 2020 Jul 23;182(2):357-371.e13. doi: 
            10.1016/j.cell.2020.05.052. Epub 2020 Jun 30.
  PUBMED    32610085
REFERENCE   2  (bases 1 to 4717)
  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: 
            "10.1016/j.cell.2020.05.052"; journalName: "Cell"; date: 
            "2020-07-23- 23"; volume: "182"; issue: "2"; pages: "357-371.e13"
COMMENT     Sequence Label: pSPLp10
FEATURES             Location/Qualifiers
     source          1..4717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..365
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(418..451)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    641..689
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     714..733
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(825..934)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1827..2279
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     2226..2247
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     primer_bind     2293..2312
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2293..2311
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3186..3320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3223..3242)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3277..3296
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3470..4261)
                     /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"
ORIGIN
        1 aattcccatg tcagccgtta agtgttcctg tgtcactcaa aattgctttg agaggctcta
       61 agggcttctc agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaagc
      121 tttaaaagcc ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta
      181 agttgctgat ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga
      241 gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa
      301 acatgagagc ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta
      361 cgtactatca acaggttgaa ctgctgatca acagatcctc tacgcggccg cggtaccata
      421 acttcgtata gcatacatta tacgaagtta tctggtttaa acgtacccgt agtggctatg
      481 gcagggcttg ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggt
      541 tggggtgggg aaaaggaaga aacgcgggcg tattggtccc aatggggtct cggtggggta
      601 tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc
      661 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
      721 cttccgtgtt tcagttagcc tcccccatct cccggtaccg catgctatgc atcagctgct
      781 agcaccatgg ctcgagatcc cgggtgatca agtcttcgtc gagtgattgt aaataaaatg
      841 taatttacag tatagtattt taattaatat acaaatgatt tgataataat tcttatttaa
      901 ctataatata ttgtgttggg ttgaattaaa ggtccgtata ctagtatcga tgattgaccc
      961 caacaaaaga tttataatta atcataatca cgaacaacaa caagtcaatg aaacaaataa
     1021 acaagttgtc gataaaacat tcataaatga cacagcaaca tacaattctt gcataataaa
     1081 aatttaaatg acatcatatt tgagaataac aaatgacatt atccctcgat tgtgttttac
     1141 aagtagaatt ctacccgtaa agcgagttta gttttgaaaa acaaatgaca tcatttgtat
     1201 aatgacatca tcccctgatt gtgttttaca agtagaattc tatccgtaaa gcgagttcag
     1261 ttttgaaaac aaatgagtca tacctaaaca cgttaataat cttctgatat cagcttatga
     1321 ctcaagttat gagccgtgtg caaaacatga gataagttta tgacatcatc cactgatcgt
     1381 gcgttacaag tagaattcta ctcgtaaagc cagttcggtt atgagccgtg tgcaaaacat
     1441 gacatcagct tatgactcat acttgattgt gttttacgcg tagaattcta ctcgtaaagc
     1501 gagttcggtt atgagccgtg tgcaaaacat gacatcagct tatgagtcat aattaatcgt
     1561 gcgttacaag tagaattcta ctcgtaaagc gagttgaagg atcatattta gttgcgttta
     1621 tgagataaga ttgaaaagcg tgtaaaatgt ttcccgcgct tggcacaact atttacaatg
     1681 cggccaagtt ataaaagatt ctaatctgat atgttttaaa acacctttgc ggcccgagtt
     1741 gtttgcgtac gtgactagcg aagaagatgt gtggaccgca gaacagatag taaaacaaaa
     1801 ccctagtatt ggagcaataa tcgatcctag aatcccaaaa caaactggtt attgtggtag
     1861 gtcatttgtt tggcagaaag aaaactcgag aaatttctct ggccgttatt cgttattctc
     1921 tcttttcttt ttgggtctct ccctctctgc actaatgctc tctcactctg tcacacagta
     1981 aacggcatac tgctctcgtt ggttcgagag agcgcgcctc gaatgttcgc gaaaagagcg
     2041 ccggagtata aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag
     2101 tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca
     2161 atctgcagta aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat
     2221 caactgcaac tactgaaatc tgccaagaag taattattga atacaagaag agaactctga
     2281 atagggaatt ggtaatacga ctcactatag ggggatcccg gtccgaagcg cgcggaattc
     2341 aaaggcctac gtcgacgagc tcactagtcg cggccgcttt cgaatctaga atgaatcgtt
     2401 tttaaaataa caaatcaatt gttttataat attcgtacga ttctttgatt atgtaataaa
     2461 atgtgatcat taggaagatt acgaaaaata taaaaaatat gagttctgtg tgtataacaa
     2521 atgctgtaaa cgccacaatt gtgtttgttg caaataaacc catgattatt tgattaaaat
     2581 tgttgttttc tttgttcata gacaatagtg tgttttgcct aaacgtgtac tgcataaact
     2641 ccatgcgagt gtatagcgag ctagtggcta acgcttgccc caccaaagta gattcgtcaa
     2701 aatcctcaat ttcatcaccc tcctccaagt ttaacatttg gccgtcggaa ttaacttcta
     2761 aagatgccac ataatctaat aaatgaaata gagattcaaa cgtggcgtca tcgtccgttt
     2821 cgaccatttc cgaaaagaac tcgggcataa actctatgat ttctctggac gtggtgttgt
     2881 cgaaactctc aaagtacgca gtcaggaacg tgcgcgacat gtcgtcggga aactcgcgcg
     2941 gaaacatgtt gttgtaaccg aacgggtccc atagcgccaa aaccaaatct gccagcgtca
     3001 atagaatgag cacgatgccg acaatggagc tggcttggat agcgattcga gttaacaagc
     3061 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
     3121 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3181 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3241 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3301 tatcttatca tgtctggatc tgatcactgc ttgagcctag aagatccggc tgctaacaaa
     3361 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactatc ataaccccta
     3421 ggagtccaag cgagctcgat atccgtcggc ttgaacgaat tgttagacat tatttgccga
     3481 ctaccttggt gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg
     3541 aggccaagcg atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct
     3601 gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt
     3661 tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc
     3721 cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat
     3781 cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat
     3841 gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga
     3901 tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat
     3961 aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct
     4021 cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg
     4081 tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc
     4141 cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct
     4201 cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca
     4261 tgatgtttaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata
     4321 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     4381 gtaccccaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     4441 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctactta ccggtttatt
     4501 gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc
     4561 tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc agctgacatt
     4621 catccggggt cagcaccgtt tctgcggact ggctttctac gtgttccgct tcctttagca
     4681 gcccttgcgc cctgagtgct tgcggcagcg tgaagct
//
