LOCUS       sequence-12265-e        6249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  AUTHORS   Dreosti E, Odermatt B, Dorostkar MM, Lagnado L
  TITLE     A genetically encoded reporter of synaptic activity in vivo.
  JOURNAL   Nat Methods. 2009 Dec . 6(12):883-9.
  PUBMED    19898484
REFERENCE   2  (bases 1 to 6249)
  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: "Nat 
            Methods. 2009 Dec . 6(12):883-9."
COMMENT     Sequence Label: CMV::ratSyGCaMP2
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1560..1571,1572..1589,1590..1592,1593..1625,1626..1628,
                     1629..1652,1653..1679,1680..1736,1737..1742,1743..2012,
                     2013..2030,2031..2465,2466..2468,2469..2471,2472..2915)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(1560..1571,1572..1589,1590..1592,1593..1625,1626..1628,
                     1629..1652,1653..1679,1680..1736,1737..1742,1743..2012,
                     2013..2030,2031..2465,2466..2468,2469..2471,2472..2915)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1659..1668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1949..1970
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2076..2097)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2337..2356)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3037..3158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3074..3093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3128..3147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3165..3620)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3302..3323)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3514..3533
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3647..3751
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3753..4110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3753..3944
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3776..3796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3961..4096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4145..4939
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4145..4939
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4199..4218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4809..4828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5127..5146)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5171..5218
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5547..6135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6036..6055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc accatggacg tggtgaatca gctggtggct gggggtcagt
      661 tccgggtggt caaggagccc cttggcttcg tgaaggtgct gcagtgggtc tttgccatct
      721 tcgcctttgc tacgtgtggc agctacaccg gggagcttcg gctgagcgtg gagtgtgcca
      781 acaagacgga gagtgccctc aacatcgaag ttgaattcga gtaccccttc aggctgcacc
      841 aagtgtactt tgatgcaccc tcctgcgtca aagggggcac taccaagatc ttcctggttg
      901 gggactactc ctcgtcggct gaattctttg tcaccgtggc tgtgtttgcc ttcctctact
      961 ccatgggggc cctggccacc tacatcttcc tgcagaacaa gtaccgagag aacaacaaag
     1021 ggcctatgat ggactttctg gctacagccg tgttcgcttt catgtggcta gttagttcat
     1081 cagcctgggc caaaggcctg tccgatgtga agatggccac ggacccagag aacattatca
     1141 aggagatgcc catgtgccgc cagacaggga acacatgcaa ggaactgagg gaccctgtga
     1201 cttcaggact caacaccaca gtggtgtttg gcttcctgaa cctggtgctc tgggttggca
     1261 acttatggtt cgtgttcaag gagacaggct gggcagcccc attcatgcgc gcacctccag
     1321 gcgccccgga aaagcaacca gcacctggcg atgcctacgg cgatgcgggc tacgggcagg
     1381 gccccggagg ctatgggccc caggactcct acgggcctca gggtggttat caacccgatt
     1441 acgggcagcc agccagcggt ggcggtggct acgggcctca gggcgactat gggcagcaag
     1501 gctatggcca acagggtgcg cccacctcct tctccaatca ggatccaccg gtcgccacca
     1561 tgcggggttc tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa
     1621 tgggtcggga tctgtacgac gatgacgata aggatctcgc caccatggtc gactcatcac
     1681 gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg agctcactcg
     1741 agaacgtcta tatcatggcc gacaagcaga agaacggcat caaggcgaac ttcaagatcc
     1801 gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca
     1861 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaaacttt
     1921 cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1981 ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagca
     2041 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2101 acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga
     2161 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2221 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2281 tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg
     2341 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2401 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2461 acaacacgcg tgaccaactg actgaagagc agatcgcaga atttaaagag gctttctccc
     2521 tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg gtgatgcggt
     2581 ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg
     2641 acggtaatgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga aaaatgaaag
     2701 acacagacag tgaagaagaa attagagaag cgttccgtgt gtttgataag gatggcaatg
     2761 gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag aagttaacag
     2821 atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt caggtaaact
     2881 acgaagagtt tgtacaaatg atgacagcga agtgagcggc cgcgactcta gatcataatc
     2941 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3001 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3061 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3121 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     3181 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3241 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3301 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3361 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3421 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3481 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3541 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3601 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3661 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3721 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3781 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3961 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4021 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4081 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4141 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4201 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4261 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4321 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4381 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4441 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4501 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4561 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4621 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4681 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4741 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4801 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4861 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4921 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4981 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5041 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5101 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5161 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5221 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5281 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5341 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     5701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6241 gccatgcat
//
