LOCUS       sequence-313165-        6853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ribosome-tagged, Cre-dependent GCaMP for soma localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6853)
  AUTHORS   Chen Y, Jang H, Spratt PWE, Kosar S, Taylor DE, Essner RA, Bai L, 
            Leib DE, Kuo TW, Lin YC, Patel M, Subkhangulova A, Kato S, Feinberg 
            EH, Bender KJ, Knight ZA, Garrison JL
  TITLE     Soma-Targeted Imaging of Neural Circuits by Ribosome Tethering.
  JOURNAL   Neuron. 2020 Jun 5. pii: S0896-6273(20)30351-2. doi: 
            10.1016/j.neuron.2020.05.005.
  PUBMED    32574560
REFERENCE   2  (bases 1 to 6853)
  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: "Neuron. 
            2020 Jun 5. pii: S0896-6273(20)30351-2. doi: 
            10.1016/j.neuron.2020.05.005."
COMMENT     Sequence Label: pyc111m
FEATURES             Location/Qualifiers
     source          1..6853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..468
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(506..539)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(590..623)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1322..1762,1763..1768,1769..2203,2204..2221,
                     2222..2491,2492..2497,2498..2554,2555..2581,2582..2605,
                     2606..2608,2609..2641,2642..2644,2645..2662,2663..2671))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1878..1897
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2137..2158
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2264..2285)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2566..2575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2668..2677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2684..2717
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2768..2801
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2826..3414
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2879..2899)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3297..3308)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3446..3922
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3645..3664)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3962..4102
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3962..4091
                     /label=AAV2 ITR
     rep_origin      4177..4632
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4264..4283)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4474..4495
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4649..4668)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4768..4790
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4828..4846)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4914..5018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5019..5087,5088..5879)
                     /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(5237..5256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6050..6638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6539..6558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6700..6829
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag gcggggtggg
       61 ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa
      121 attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca
      181 ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc
      241 cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc
      301 ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc
      361 gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct
      421 cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac
      481 cggatcctct agagtcgact ccggaataac ttcgtatagg atactttata cgaagttatg
      541 cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa taacttcgta
      601 tagcatacat tatacgaagt tatggcgcgc cctaatacag acgctggggc ttgcccatgg
      661 tgctcttgat gtacagagcc cgcacgtttt gccagttttt cttaagcaag gacaccaaga
      721 aattgacagc cagatgaatg ttgtagacta gctcatcatc ggtcatcttc acgtggccaa
      781 cagcgacggc caaacacagc accttcttca tctggaactt gattgtcgat ttcacctcat
      841 ccactttggc caccatgttt tcattgtgtg tcagcaggga ggggaacttg ccagccttgt
      901 ttaggcctgg gcccaggata cgtgggatct gcttaatcag agactcagag gccaaaaagg
      961 catcgtactt cttagccagc ttcttgacca acttcttgtt tttgttaagt ttcttgagcg
     1021 cctcgatgtc catgtggggg atatccacgg ccttggcttc atcacagtgc tgctggtccc
     1081 ccagaacgca caccgagaac ttggggcgtg gggtggactt gagcctgacg gtgcccgaga
     1141 aacgtttgtc cttctgaggg tcgtagttct tcaggctgat ctgcagctcc accgtctcca
     1201 gaaacttgcg gcgcttgcgc tggttcccgt gcaggacttc ccgcaccgcc tcgtacaggg
     1261 tgtcgcgtga gactttgctg ctgaactcga agcttgagct tgagatctga gtccggcctg
     1321 acttcgctgt catcatttgt acaaactctt cgtagtttac ctgaccatcc ccatcgatgt
     1381 ctgcttccct gatcatttca tcaacctctt catctgttaa cttctctcca aggtttgtca
     1441 tcacgtggcg aagctctgct gcactgatgt agccattgcc atccttatca aacacaccga
     1501 acgcttctct aatttcttct tccgtgtccc tgtagctccc ttttcttgcc atcattgtca
     1561 ggaactcagg gaagtcgatt gtgccgtcac cgtcggcatc tacttcattg atcatgtcct
     1621 gcagctctgc ttctgtgggg ttctgcccca gagaccgcat caccgtcccc agctccttgg
     1681 ttgttattgt cccatccccg tccttgtcaa atagggagaa agcctcttta aattctgcga
     1741 tctgctcttc agtcagttgg tccggcaggt tgtactccag cttgtgcccc aggatgttgc
     1801 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     1861 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     1921 ggatgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     1981 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     2041 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     2101 caccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     2161 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catgctccct ccggtaccgc
     2221 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2281 ccatgtgatc gcgcttctcg ttggggtctt tcgaaagttt ggactgcacg ctcaggtagt
     2341 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtagg
     2401 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttcgcc ttgatgccgt
     2461 tcttctgctt gtcggccttg atatagacgt tctcgagtga gctcagccga cctatagctc
     2521 tgactgcgtg acctgtctta ttccacttac gacgtgatga gtcgaccatg gtggcgagat
     2581 ccttatcgtc atcgtcgtac agatcccgac ccatttgctg tccaccagtc atgctagcca
     2641 taccatgatg atgatgatga tgagaaccca tggtggcgct agcataactt cgtataaagt
     2701 atcctatacg aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt
     2761 gcaaagaata acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta
     2821 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2881 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2941 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3001 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3061 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3121 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3181 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3241 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3301 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3361 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3421 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     3481 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3541 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3601 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3661 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3721 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3781 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3841 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3901 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     3961 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4021 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4081 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4141 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4201 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4261 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4321 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4381 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4441 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4501 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4561 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4621 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4681 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4741 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4801 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4861 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4921 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4981 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5041 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5101 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5161 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5221 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5281 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5341 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5401 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5461 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5521 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5581 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5641 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5701 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5761 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5821 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5881 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5941 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6001 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6061 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6121 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6181 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6241 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6301 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6361 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6421 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6481 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6541 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6601 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6661 tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc
     6721 gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     6781 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg
     6841 cgtgtgtcta gac
//
