LOCUS       sequence-196803-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GCaMP6s in Hydra.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  AUTHORS   Dupre C, Yuste R
  TITLE     Non-overlapping Neural Networks in Hydra vulgaris.
  JOURNAL   Curr Biol. 2017 Apr 24;27(8):1085-1097. doi: 
            10.1016/j.cub.2017.02.049. Epub 2017 Mar 30.
  PUBMED    28366745
REFERENCE   2  (bases 1 to 5756)
  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: 
            "10.1016/j.cub.2017.02.049"; journalName: "Curr Biol"; date: 
            "2017-04-24- 24"; volume: "27"; issue: "8"; pages: "1085-1097"
COMMENT     Sequence Label: HyGCaMP6s
FEATURES             Location/Qualifiers
     source          1..5756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(103..894,895..963))
                     /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     726..745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(964..1068)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1094..1549)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1231..1252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1443..1462
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1676..1698
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1690..1707
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1691..1707
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1717..1736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1717..1735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2814..2831
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(2871..2879,2880..2894)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2922..2978
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(4659..4679)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4659..4677)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4698..4714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4698..4714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4711..4733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4722..4738
                     /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)."
     promoter        complement(join(4746..4752,4753..4770,4771..4776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4791..4812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4929..4946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5100..5688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5180..5199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
       61 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1081 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     1141 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     1201 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1261 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     1321 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     1381 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     1441 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     1501 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     1561 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     1621 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     1681 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
     1741 ttggttttaa gccgagatgc aggattctga atgaaaaaga aaaaaagaag tctcggtaga
     1801 gtaaaagtga tcggtttgca actgtaaaat ttattgaagt accaataatt ttatttaaaa
     1861 taaaactgaa atataaagtt aaagttgctg ttctataagt ttaccgaatt ttaaaaccat
     1921 tgtaacgcta gagtaatatt tgagtctact aagttagtcc ccgcactttt taatcaagca
     1981 ataaataccc aaactttgct tattcaaatc aataaaccaa tatatctctt aaaataaagt
     2041 aaaaacttct gaaattctat aaaaaaaaat ttaatttcga aatatcaaat gtaacttcaa
     2101 caccgcacta ttttctttta aacaactgat atagtaatta cttctcaaaa acgttatctc
     2161 aaggtttgtg atgtacttaa aaccactcct attttgttac gcgtttaaaa aagcaaacat
     2221 aagttggttt ctattgatga atgagaacat atttcattta aagttaaaat cctaccagtg
     2281 gtttcactgt acgtaaacac cgtcaaaaaa acaggaacgt ttttaaagat taataattga
     2341 agtaaaaaaa atttaatacc gggggttaaa aaaatctttt aaaataatta taaatatata
     2401 tattaaaatt tataaatttt taaacacatt taaaatatat attaagtata ataaaagtaa
     2461 tattataaaa aaaaatttaa ttttataatt atttttatta aatttataaa taataggtaa
     2521 aacttacata tccgttttat tttttcttaa taaaataacg cgtgcaaatt tttgtccata
     2581 taaagacctt ttcgaacaat aacttttttg cttagccgtt ttttttctta tatggtcaaa
     2641 aaagcgctca agcgattcac cataaaaagc gcaattagtt cagcgttcgt tattcagaag
     2701 cttcagcttt gcttgatact cagctcttct ctttttaaac aaaacactta atcaaaatgg
     2761 ccgatgatga agttgccgcc ctcgctgcag ccccggtaga aaaaatgggt agtcatcacc
     2821 atcatcacca cggaatggca tcaatgacag gaggacagca aatgggtcgt gatttgtatg
     2881 acgacgatga taaagatttg gctactatgg ttgattcaag ccgccgtaaa tggaataaga
     2941 ctggtcatgc tgtacgagct attggaaggt taagttccct tgaaaatgtt tatataaaag
     3001 cagataagca gaaaaatggt atcaaagcta attttcatat acgacataac attgaagatg
     3061 gtggtgttca acttgcatat cactatcaac agaatactcc tattggcgac ggacctgttt
     3121 tacttcctga taatcattat ctatctgtcc aatcaaaact gtctaaagac ccaaacgaaa
     3181 aacgtgatca tatggttctt ttggaatttg tgacggctgc cggtattaca ttagggatgg
     3241 atgagcttta caaagggggt accggtggta gtatggtttc gaaaggagaa gagttgttta
     3301 ctggagtagt accaattctg gttgaattag acggagatgt taatggtcat aaattttcag
     3361 tttctggaga aggggaagga gatgctacat atggaaaact tacgcttaag ttcatatgca
     3421 ccacaggaaa attgcctgtt ccatggccca ccttggttac aacattaaca tatggtgtcc
     3481 aatgttttag tagatatcca gatcatatga aacaacatga ctttttcaaa tctgcaatgc
     3541 cagaaggcta tattcaagag agaactatat tttttaagga tgatggaaac tataagacaa
     3601 gagctgaagt gaaatttgaa ggagatactt tagtgaatcg catagaactt aaaggtatag
     3661 actttaaaga ggatggcaat attttaggtc acaaattaga gtacaatcta cctgatcaac
     3721 taacagagga acagattgca gagttcaaag aggcattttc actttttgat aaagatggcg
     3781 atggaacaat cactactaag gaattaggta ctgtaatgag atcacttggt caaaatccta
     3841 ctgaagcgga attacaagat atgattaacg aagtagatgc agatggggat ggaactatag
     3901 actttccgga atttttaaca atgatggcac gaaaaatgaa ataccgtgat actgaagagg
     3961 aaattagaga agcatttggt gtatttgaca aagatggaaa cggttacatt tctgctgctg
     4021 aactaagaca tgttatgaca aacttaggag aaaaactgac tgatgaagag gtcgacgaaa
     4081 tgataagaga agccgatatt gatggagatg gtcaagttaa ttacgaagag tttgttcaaa
     4141 tgatgacagc aaagtaagaa ttcacaattc gattatattt atactggact atttttacat
     4201 ctgttcggtt attttcacat ttatttttct atatatatct tataaacgtt ttaaaaccca
     4261 tgtaattttt gttaagctgt aatataaaag acgtcctaac aaacttcttt tattactgaa
     4321 tttcctttaa ttataataaa taacaagttt taaaataaat tcaggcaatt aaggcgctcc
     4381 tgaggtacta aaattaatgt aaacatttaa aattaacttg gatggtctta agtactgtac
     4441 tcgtgatttt gttatacttt attattagaa aagtcgtcta ttaacttttt gttccttaat
     4501 ttacttgatt aaattgtcgc ttaatttatc aaatcaggtt ttgcgcgtta ttttagagaa
     4561 aaacttatta gaaaaatgaa taagcaaagt ttaggctaac atgttttttt attattttaa
     4621 atagttcaag tcaatgacgt ataaaccagc ttttgttccc tttagtgagg gttaattgcg
     4681 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4741 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4801 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4861 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4921 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4981 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5521 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gatttt
//
