LOCUS       sequence-313189-        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: pyc111s
FEATURES             Location/Qualifiers
     source          1..6853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(486..519)
                     /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(570..603)
                     /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(1302..1742,1743..1748,1749..2183,2184..2201,
                     2202..2471,2472..2477,2478..2534,2535..2561,2562..2585,
                     2586..2588,2589..2621,2622..2624,2625..2642,2643..2651))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1858..1877
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2117..2138
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2244..2265)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2546..2555
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2648..2657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2664..2697
                     /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    2748..2781
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2806..3394
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2859..2879)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3277..3288)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3426..3902
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3625..3644)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3942..4082
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3942..4071
                     /label=AAV2 ITR
     rep_origin      4157..4612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4244..4263)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4454..4475
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4629..4648)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4748..4770
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4808..4826)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4894..4998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4999..5067,5068..5859)
                     /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(5217..5236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6030..6618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6519..6538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6680..6809
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatcctct agagtcgact
      481 ccggaataac ttcgtatagg atactttata cgaagttatg cagaatggta gctggattgt
      541 agctgctatt agcaatatga aacctcttaa taacttcgta tagcatacat tatacgaagt
      601 tatggcgcgc cctaatacag acgctggggc ttgcccatgg tgctcttgat gtacagagcc
      661 cgcacgtttt gccagttttt cttaagcaag gacaccaaga aattgacagc cagatgaatg
      721 ttgtagacta gctcatcatc ggtcatcttc acgtggccaa cagcgacggc caaacacagc
      781 accttcttca tctggaactt gattgtcgat ttcacctcat ccactttggc caccatgttt
      841 tcattgtgtg tcagcaggga ggggaacttg ccagccttgt ttaggcctgg gcccaggata
      901 cgtgggatct gcttaatcag agactcagag gccaaaaagg catcgtactt cttagccagc
      961 ttcttgacca acttcttgtt tttgttaagt ttcttgagcg cctcgatgtc catgtggggg
     1021 atatccacgg ccttggcttc atcacagtgc tgctggtccc ccagaacgca caccgagaac
     1081 ttggggcgtg gggtggactt gagcctgacg gtgcccgaga aacgtttgtc cttctgaggg
     1141 tcgtagttct tcaggctgat ctgcagctcc accgtctcca gaaacttgcg gcgcttgcgc
     1201 tggttcccgt gcaggacttc ccgcaccgcc tcgtacaggg tgtcgcgtga gactttgctg
     1261 ctgaactcga agcttgagct tgagatctga gtccggcctg acttcgctgt catcatttgt
     1321 acaaactctt cgtagtttac ctgaccatcc ccatcgatgt ctgcttccct gatcatttca
     1381 tcaacctctt catctgttaa cttctctcca aggtttgtca tcacgtggcg aagctctgct
     1441 gcactgatgt agccattgcc atccttatca aacacaccga acgcttctct aatttcttct
     1501 tccgtgtccc tgtatttcat ttttcttgcc atcattgtca ggaactcagg gaagtcgatt
     1561 gtgccgtcac cgtcggcatc tacttcattg atcatgtcct gcagctctgc ttctgtgggg
     1621 ttctgcccca gagaccgcat caccgtcccc agctccttgg ttgttattgt cccatccccg
     1681 tccttgtcaa atagggagaa agcctcttta aattctgcga tctgctcttc agtcagttgg
     1741 tccggcaggt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     1801 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     1861 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggatgtagcc ttcgggcatg
     1921 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     1981 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     2041 atgaacttca gggtcagctt gccgtaggtg gcatcgccct caccctcgcc ggacacgctg
     2101 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     2161 agctcctcgc ccttgctcac catgctccct ccggtaccgc ccttgtacag ctcgtccatg
     2221 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     2281 ttggggtctt tcgaaagttt ggactgcacg ctcaggtagt ggttgtcggg cagcagcacg
     2341 gggccgtcgc cgatgggggt gttctgctgg tagtggtagg cgagctgcac gccgccgtcc
     2401 tcgatgttgt ggcggatgtg gaagttcgcc ttgatgccgt tcttctgctt gtcggccttg
     2461 atatagacgt tctcgagtga gctcagccga cctatagctc tgactgcgtg acctgtctta
     2521 ttccacttac gacgtgatga gtcgaccatg gtggcgagat ccttatcgtc atcgtcgtac
     2581 agatcccgac ccatttgctg tccaccagtc atgctagcca taccatgatg atgatgatga
     2641 tgagaaccca tggtggcgct agcataactt cgtataaagt atcctatacg aagttatttg
     2701 ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata acttcgtata
     2761 atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca acctctggat
     2821 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2881 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2941 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3001 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3061 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3121 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3181 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     3241 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3301 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3361 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     3421 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     3481 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     3541 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     3601 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     3661 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     3721 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     3781 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     3841 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     3901 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3961 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4021 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     4081 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4141 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4201 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     4261 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4321 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     4381 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4441 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     4501 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     4561 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4621 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4681 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4741 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4801 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4861 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4921 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4981 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5041 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5101 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5161 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5221 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5281 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5341 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5401 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5461 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5521 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5581 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5641 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5701 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5761 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5821 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5881 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5941 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6001 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6661 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     6721 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     6781 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtgtgtcta gactgcagag
     6841 ggccctgcgt atg
//
