LOCUS       sequence-130794-        5826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Y-GECO1 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5826)
  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 5826)
  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-Y-GECO1
FEATURES             Location/Qualifiers
     source          1..5826
                     /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)"
     CDS             1073..1129
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             2083..2097
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    2336..2924
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2389..2409)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2807..2818)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2927..2943)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2928..2944)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2973..3094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2984..3003)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3038..3057
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3256..3385
                     /label=AAV2 ITR
     primer_bind     complement(3404..3421)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3404..3420)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3413..3435)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3562..4017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3649..3668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3859..3880
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4043..4147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4148..4216,4217..5008)
                     /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(4366..4385)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5179..5767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5668..5687
                     /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 atccaccatg gtcgactcat cacgtcgtaa
     1081 gtggaataag acaggtcacg cagtcagagc tataggccgg ctgagctcac tcgagggtta
     1141 tagtaccgag aaaattgtgc ctgttccaaa gcaagggata ttaaaaggaa aggtcagcat
     1201 gtacctattg ctgaaggatg gcggtcgtta tcattgccag tttgatacgg tatataaagc
     1261 taaatcagtg ccctccaaga tgccaggacg gcattttata cagcataagc tccttaggga
     1321 ggaccgctct gatgctaaaa accataaatg gcaattaaca gaacatgcaa tcgcaggcat
     1381 ggacgagctg tacaagggag gtacaggcgg gagtatggtg agcaagggcg aggggcaatc
     1441 gaagcatggg ctaaaggagg agatgacggt gaagtaccac atggaaggtt gcgttaatgg
     1501 tcacaaattt gtcattactg gagagggcat tggaaaccct tttaagggta agcaaactgc
     1561 aaatttgtgt gtgatagaag gaggcccgct gccgttctcg gaggacattt taagtccggg
     1621 ttttaaatat ggtgaccgga ttttcacaga gtacccacag gatattgtag attacttcaa
     1681 gaactcatgt ccagagggct atacgtggga aaggagcttc ttctatgagg acggagcggt
     1741 ctgtcgatgc aacgcggaca taacagtctc tgaaaaggag aactgcatct atcacaaaag
     1801 tatcttctat ggggtgaatt ttcccgccga cggccccgta atgaagaaaa tgaccactaa
     1861 ttgggaagct gacacgcgtg accaactgac tgaagagcag atcgcagaat ttaaagaggc
     1921 tttctcccta tttgacaagg acggggatgg gacgatgaca accgaggagc tggggacggt
     1981 gctgcggtct ctggggcaga acccctcaga agcagagctg cgggacatga tcaatgaagt
     2041 agatgccgac ggtgacggca caatcggctt ccctgagttc ctgacgatga tgacaagaaa
     2101 aatgaatgac acggacagtg aagaggaaat tagagaagcg ttccgtgtgt ttgataagga
     2161 cggcaatggc tacatcggcg cagcagagct tcgccacgtg atgacaaacc ttggagagaa
     2221 gttaacagat gaagagtttg atgaaatgat cagggtagca gactccgacg gggatggtca
     2281 ggtaaactac gaagagtttg tacaaatgat gacagcaaag tgaaagctta tcgataatca
     2341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2461 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2521 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2581 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2641 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2701 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     2761 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2821 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2881 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     2941 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3001 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3061 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3121 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3181 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     3241 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3301 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3361 cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac
     3421 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3481 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3541 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3601 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3661 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3721 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3781 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3841 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3901 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3961 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     4021 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4081 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4141 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4201 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4261 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4321 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4381 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4441 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4501 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4561 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4621 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4681 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4741 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4801 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4861 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4921 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4981 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5041 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5101 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5161 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5221 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5281 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5341 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5401 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5461 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5521 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5581 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5641 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5701 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5761 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5821 tcacat
//
