LOCUS       sequence-308471-        7238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Independent expression of QuasAr and GCaMP under human synapsin 1 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7238)
  AUTHORS   Cho IH, Panzera LC, Chin M, Alpizar SA, Olveda GE, Hill RA, Hoppa MB
  TITLE     The potassium channel subunit K(v)beta1 serves as a major control 
            point for synaptic facilitation.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Nov 24;117(47):29937-29947. doi: 
            10.1073/pnas.2000790117. Epub 2020 Nov 9.
  PUBMED    33168717
REFERENCE   2  (bases 1 to 7238)
  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: 
            "10.1073/pnas.2000790117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-11-24- 24"; volume: "117"; issue: "47"; pages: 
            "29937-29947"
COMMENT     Sequence Label: QuasAr-P2A-GCaMP
FEATURES             Location/Qualifiers
     source          1..7238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        212..659
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      694..703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             700..1458
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     CDS             1465..2172
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTAGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             2254..2310
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2311..2319,2320..2337,2338..2340,2341..2373,2374..2376,
                     2377..2400,2401..2427,2428..2484,2485..2490,2491..2760,
                     2761..2778,2779..3213,3214..3219,3220..3663)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      2407..2416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2697..2718
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2824..2845)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3085..3104)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3676..4264
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3729..3749)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4147..4158)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4267..4283)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4268..4284)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4313..4434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4324..4343)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4378..4397
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4596..4725
                     /label=AAV2 ITR
     rep_origin      complement(4962..5550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5042..5061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5721..6512,6513..6581))
                     /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     6344..6363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6582..6686)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6712..7167)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6849..6870)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7061..7080
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga
      241 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
      301 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
      361 gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg
      421 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
      481 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
      541 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
      601 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt
      661 cgaattcaag ctgctagcaa ggatcctcta gacgccacca tggtaagtat cgctctgcag
      721 gctggttacg acctactggg tgacggcaga cctgaaaccc tgtggctggg catcggcact
      781 ctgctgatgc tgattggaac cttctacttt ctggtccgcg gatggggagt caccgataag
      841 gatgcccggg aatattacgc tgtgactatc ctggtgtccg gaatcgcatc cgccgcatat
      901 ctgtctatgt tctttggtat cgggcttact gaggtgtccg tcgggggcga aatgttggat
      961 atctattatg ccaggtacgc ccagtggctg tttaccaccc cacttctgct gctgcacctg
     1021 gcccttctcg ctaaggtgga tcgggtgacc atcggcaccc tggtgggtgt ggacgccctg
     1081 atgatcgtca ctggcctcat cggagccttg agccacacgg ccatagccag atacagttgg
     1141 tggttgttct ctacaatttg catgatagtg gtgctctatg ttctggctac atccctgcga
     1201 tctgctgcaa aggagcgggg ccccgaggtg gcatctacct ttaacaccct gacagctctg
     1261 gtcttggtgc tgtggaccgc ttaccctatc ctgtggatca taggcactga gggcgctggc
     1321 gtggtgggcc tgggcatcga aactctgctg tttatggtgt tggacgtgac tgccaaggtc
     1381 ggctttggct ttatcctgtt gagatcccgg gctattctgg gcgacaccga ggcaccagaa
     1441 cccagtgccg gtgccgatac gttgatggtg agcaagggcg aggagtataa catggccatc
     1501 atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg ccacgagttc
     1561 gagatcgagg gcgagggcga gggccgcccc tacgagggct ttcagaccgc taagctgaag
     1621 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca tttcaccgcc
     1681 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttcaa gctgtccttc
     1741 cccgagggct tcaagtggga gcgcgtgatg aactacgagg acggcggcgt ggtgaccgtg
     1801 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1861 aacttcccct ccgacggccc cgtgatgcag aagaagacca tgggctggga ggcctcctcc
     1921 gagcggatgt accccgagga cggtgccctg aagggcaaga tcaagatgag gctgaagctg
     1981 aaggacggcg gccactacac ctccgaggtc aagaccacct acaaggccaa gaagcccgtg
     2041 cagctgcccg gcgcctacat cgtcgacatc aagttggaca tcacctccca caacgaggac
     2101 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2161 gagctgtaca agagtagaat cacaagcgaa ggcgagtaca tccccctgga tcaaatagac
     2221 ataaatgtag gtggattttg ttatgagaat gaagctacta acttcagcct gctgaagcag
     2281 gctggagacg tggaggagaa ccctggacct atgggttctc atcatcatca tcatcatggt
     2341 atggctagca tgactggtgg acagcaaatg ggtcgggatc tgtacgacga tgacgataag
     2401 gatctcgcca ccatggtcga ctcatcacgt cgtaagtgga ataagacagg tcacgcagtc
     2461 agagctatag gtcggctgag ctcactcgag aacgtctata tcaaggccga caagcagaag
     2521 aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     2581 gcctaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2641 cactacctga gcgtgcagtc caaactttcg aaagacccca acgagaagcg cgatcacatg
     2701 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2761 ggcggtaccg gagggagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2821 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt
     2881 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2941 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3001 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc
     3061 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3121 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3181 ggcaacatcc tggggcacaa gctggagtac aacctgccgg accaactgac tgaagagcag
     3241 atcgcagaat ttaaagagga attctcccta tttgacaagg acggggatgg gacaataaca
     3301 accaaggagc tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg
     3361 caggacatga tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc
     3421 ctgacaatga tggcaagaaa aatgaaatac agggacacgg aagaagaaat tagagaagcg
     3481 ttcggtgtgt ttgataagga tggcaatggc tacatcagtg cagcagagct tcgccacgtg
     3541 atgacaaacc ttggagagaa gttaacagat gaagaggttg atgaaatgat cagggaagca
     3601 gacatcgatg gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag
     3661 tgaaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3721 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3781 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3841 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3901 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3961 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4021 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     4081 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4141 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4201 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4261 ccgcatcgat accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac
     4321 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4381 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4441 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     4501 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     4561 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     4621 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4681 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     4741 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6541 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6601 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6661 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     6721 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6781 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6841 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6901 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6961 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     7021 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     7081 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     7141 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     7201 aagggcgatc ggtgcgggcc tcttcgctat tacgccag
//
