LOCUS       sequence-89440-e        5895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the membrane targeted Y-GECO1 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5895)
  AUTHORS   Zhao Y, Abdelfattah AS, Zhao Y, Ruangkittisakul A, Ballanyi K, 
            Campbell RE, Harrison DJ
  TITLE     Microfluidic cell sorter-aided directed evolution of a protein-based
            calcium ion indicator with an inverted fluorescent response.
  JOURNAL   Integr Biol (Camb). 2014 May 19.
  PUBMED    24840546
REFERENCE   2  (bases 1 to 5895)
  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: "Integr Biol
            (Camb). 2014 May 19."
COMMENT     Sequence Label: AAV-hSyn-Lyn-Y-GECO1
FEATURES             Location/Qualifiers
     source          1..5895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    229..250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(265..270,271..288,289..295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    303..319
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     308..330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     327..343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     327..343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   381..510
                     /label=AAV2 ITR
     promoter        592..1038
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1055..1064
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2152..2166
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    2405..2993
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2458..2478)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2876..2887)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2996..3012)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2997..3013)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3042..3163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3053..3072)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3107..3126
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3325..3454
                     /label=AAV2 ITR
     primer_bind     complement(3473..3490)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3473..3489)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3482..3504)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3631..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3718..3737)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3928..3949
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4112..4216
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4217..4285,4286..5077)
                     /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(4435..4454)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5248..5836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5737..5756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      181 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      241 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      301 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt
      361 taattaaggc cttaattagg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
      421 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      481 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
      541 tatctacgta gccatgctct aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt
      601 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccaccca
      661 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
      721 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
      781 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca
      841 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc
      901 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg
      961 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg
     1021 tcgtgcctga gagcgcagtc gaattcgggg atccgccacc atgggatgta taaagagtaa
     1081 aagaaaagat aacttaaatg atgacgaaga tgataaggat ctcgccacaa tggccgactc
     1141 atcacgtcgt aagtggaata agacacacgc agtcagagct ataggccggc tgagctcact
     1201 cgagggttat agtaccgaga aaattgtgcc tgttccaaag caagggatat taaaaggaaa
     1261 ggtcagcatg tacctattgc tgaaggatgg cggtcgttat cattgccagt ttgatacggt
     1321 atataaagct aaatcagtgc cctccaagat gccaggacgg cattttatac agcataagct
     1381 ccttagggag gaccgctctg atgctaaaaa ccataaatgg caattaacag aacatgcaat
     1441 cgcaggcatg gacgagctgt acaagggagg tacaggcggg agtatggtga gcaagggcga
     1501 ggggcaatcg aagcatgggc taaaggagga gatgacggtg aagtaccaca tggaaggttg
     1561 cgttaatggt cacaaatttg tcattactgg agagggcatt ggaaaccctt ttaagggtaa
     1621 gcaaactgca aatttgtgtg tgatagaagg aggcccgctg ccgttctcgg aggacatttt
     1681 aagtccgggt tttaaatatg gtgaccggat tttcacagag tacccacagg atattgtaga
     1741 ttacttcaag aactcatgtc cagagggcta tacgtgggaa aggagcttct tctatgagga
     1801 cggagcggtc tgtcgatgca acgcggacat aacagtctct gaaaaggaga actgcatcta
     1861 tcacaaaagt atcttctatg gggtgaattt tcccgccgac ggccccgtaa tgaagaaaat
     1921 gaccactaat tgggaagctg acacgcgtga ccaactgact gaagagcaga tcgcagaatt
     1981 taaagaggct ttctccctat ttgacaagga cggggatggg acgatgacaa ccgaggagct
     2041 ggggacggtg ctgcggtctc tggggcagaa cccctcagaa gcagagctgc gggacatgat
     2101 caatgaagta gatgccgacg gtgacggcac aatcggcttc cctgagttcc tgacgatgat
     2161 gacaagaaaa atgaatgaca cggacagtga agaggaaatt agagaagcgt tccgtgtgtt
     2221 tgataaggac ggcaatggct acatcggcgc agcagagctt cgccacgtga tgacaaacct
     2281 tggagagaag ttaacagatg aagagtttga tgaaatgatc agggtagcag actccgacgg
     2341 ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcaaagt gaaagcttat
     2401 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     2821 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3001 ccgtcgacct cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3061 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3121 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3181 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3241 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     3301 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     3361 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3421 cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat tcactggccg
     3481 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3541 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3601 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3661 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3721 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3781 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3841 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3901 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3961 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4021 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4081 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4141 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4201 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4261 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4321 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4381 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4441 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4501 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4561 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4621 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4681 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4741 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4801 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4861 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4921 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4981 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5041 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5101 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5161 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5221 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5281 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5341 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5401 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5461 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5521 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5581 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5641 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5701 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5761 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5821 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5881 gccttttgct cacat
//
