LOCUS       sequence-335760-        7351 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 7351)
  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 7351)
  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: pFp10
FEATURES             Location/Qualifiers
     source          1..7351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4048..4081)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4271..4319
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4344..4363
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4455..4564)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5457..5909
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     5856..5877
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     primer_bind     5923..5942
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5923..5941
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6816..6950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6853..6872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6907..6926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7104..7269
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttt
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cagctttgtt tgtactatca acaggttgaa ctgctgatca
     4021 acagatcctc tacgcggccg cggtaccata acttcgtata gcatacatta tacgaagtta
     4081 tctggtttaa acgtacccgt agtggctatg gcagggcttg ccgccccgac gttggctgcg
     4141 agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga aacgcgggcg
     4201 tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     4261 ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt ttattgccgt
     4321 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tcccccatct
     4381 cccggtaccg catgctatgc atcagctgct agcaccatgg ctcgagatcc cgggtgatca
     4441 agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt taattaatat
     4501 acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg ttgaattaaa
     4561 ggtccgtata ctagtatcga tgattgaccc caacaaaaga tttataatta atcataatca
     4621 cgaacaacaa caagtcaatg aaacaaataa acaagttgtc gataaaacat tcataaatga
     4681 cacagcaaca tacaattctt gcataataaa aatttaaatg acatcatatt tgagaataac
     4741 aaatgacatt atccctcgat tgtgttttac aagtagaatt ctacccgtaa agcgagttta
     4801 gttttgaaaa acaaatgaca tcatttgtat aatgacatca tcccctgatt gtgttttaca
     4861 agtagaattc tatccgtaaa gcgagttcag ttttgaaaac aaatgagtca tacctaaaca
     4921 cgttaataat cttctgatat cagcttatga ctcaagttat gagccgtgtg caaaacatga
     4981 gataagttta tgacatcatc cactgatcgt gcgttacaag tagaattcta ctcgtaaagc
     5041 cagttcggtt atgagccgtg tgcaaaacat gacatcagct tatgactcat acttgattgt
     5101 gttttacgcg tagaattcta ctcgtaaagc gagttcggtt atgagccgtg tgcaaaacat
     5161 gacatcagct tatgagtcat aattaatcgt gcgttacaag tagaattcta ctcgtaaagc
     5221 gagttgaagg atcatattta gttgcgttta tgagataaga ttgaaaagcg tgtaaaatgt
     5281 ttcccgcgct tggcacaact atttacaatg cggccaagtt ataaaagatt ctaatctgat
     5341 atgttttaaa acacctttgc ggcccgagtt gtttgcgtac gtgactagcg aagaagatgt
     5401 gtggaccgca gaacagatag taaaacaaaa ccctagtatt ggagcaataa tcgatcctag
     5461 aatcccaaaa caaactggtt attgtggtag gtcatttgtt tggcagaaag aaaactcgag
     5521 aaatttctct ggccgttatt cgttattctc tcttttcttt ttgggtctct ccctctctgc
     5581 actaatgctc tctcactctg tcacacagta aacggcatac tgctctcgtt ggttcgagag
     5641 agcgcgcctc gaatgttcgc gaaaagagcg ccggagtata aatagaggcg cttcgtctac
     5701 ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa
     5761 ataaacaagc gcagctgaac aagctaaaca atctgcagta aagtgcaagt taaagtgaat
     5821 caattaaaag taaccagcaa ccaagtaaat caactgcaac tactgaaatc tgccaagaag
     5881 taattattga atacaagaag agaactctga atagggaatt ggtaatacga ctcactatag
     5941 ggggatcccg gtccgaagcg cgcggaattc aaaggcctac gtcgacgagc tcactagtcg
     6001 cggccgcttt cgaatctaga atgaatcgtt tttaaaataa caaatcaatt gttttataat
     6061 attcgtacga ttctttgatt atgtaataaa atgtgatcat taggaagatt acgaaaaata
     6121 taaaaaatat gagttctgtg tgtataacaa atgctgtaaa cgccacaatt gtgtttgttg
     6181 caaataaacc catgattatt tgattaaaat tgttgttttc tttgttcata gacaatagtg
     6241 tgttttgcct aaacgtgtac tgcataaact ccatgcgagt gtatagcgag ctagtggcta
     6301 acgcttgccc caccaaagta gattcgtcaa aatcctcaat ttcatcaccc tcctccaagt
     6361 ttaacatttg gccgtcggaa ttaacttcta aagatgccac ataatctaat aaatgaaata
     6421 gagattcaaa cgtggcgtca tcgtccgttt cgaccatttc cgaaaagaac tcgggcataa
     6481 actctatgat ttctctggac gtggtgttgt cgaaactctc aaagtacgca gtcaggaacg
     6541 tgcgcgacat gtcgtcggga aactcgcgcg gaaacatgtt gttgtaaccg aacgggtccc
     6601 atagcgccaa aaccaaatct gccagcgtca atagaatgag cacgatgccg acaatggagc
     6661 tggcttggat agcgattcga gttaacaagc ttgtcgagaa gtactagagg atcataatca
     6721 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     6781 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     6841 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6901 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc
     6961 ttgagcctag aagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     7021 accgctgagc aataactatc ataacctaac cccttggggc ctctaaacgg gtcttgaggg
     7081 gttttttgac cctaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt
     7141 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     7201 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     7261 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc
     7321 catgtgacaa accgtcatct tcggctactt t
//
