LOCUS       sequence-196624-        5757 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 5757)
  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 5757)
  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..5757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     115..132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     116..132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     142..161
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        142..160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1240..1257
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(1297..1305,1306..1320)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1348..1404
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(3085..3105)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3085..3103)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3124..3140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3124..3140)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3137..3159)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3148..3164
                     /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(3172..3178,3179..3196,3197..3202))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3217..3238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3355..3372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3526..4114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3606..3625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4285..5076,5077..5145))
                     /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     4908..4927
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5146..5250)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5276..5731)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5413..5434)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5625..5644
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
       61 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
      121 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggt tttaagccga
      181 gatgcaggat tctgaatgaa aaagaaaaaa agaagtctcg gtagagtaaa agtgatcggt
      241 ttgcaactgt aaaatttatt gaagtaccaa taattttatt taaaataaaa ctgaaatata
      301 aagttaaagt tgctgttcta taagtttact cgaattttaa aaccattgta acgctagagt
      361 aatatttgag tctactaagt tagtccccgc actttttaat caagcaataa atacccaaac
      421 tttgcttatt caaatcaata aaccaatata tctcttaaaa taaagtaaaa acttctgaaa
      481 ttctataaaa aaaaatttaa tttcgaaata tcaaatgtaa cttcaacacc gcactatttt
      541 cttttaaaca actgatatag taattacttc tcaaaaacgt tatctcaagg tttgtgatgt
      601 acttaaaacc actcctattt tgttacgcgt ttaaaaaagc aaacataagt tggtttctat
      661 tgatgaatga gaacatattt catttaaagt taaaatccta ccagtggttt cactgtacgt
      721 aaacaccgtc aaaaaaacag gaacgttttt aaagattaat aattgaagta aaaaaaattt
      781 aataccgggg gttaaaaaaa tcttttaaaa taattataaa tatatatatt aaaatttata
      841 aatttttaaa cacatttaaa atatatatta agtataataa aagtaatatt ataaaaaaaa
      901 atttaatttt ataattattt ttattaaatt tataaataat aggtaaaact tacatatccg
      961 ttttattttt tcttaataaa ataacgcgtg caaatttttg tccatataaa gaccttttcg
     1021 aacaataact tttttgctta gccgtttttt ttcttatatg gtcaaaaaag cgctcaagcg
     1081 attcaccata aaaagcgcaa ttagttcagc gttcgttatt cagaagcttc agctttgctt
     1141 gatactcagc tcttctcttt ttaaacaaaa cacttaatca aaatggccga tgatgaagtt
     1201 gccgccctcg ctgcagcccc ggtagaaaaa atgggtagtc atcaccatca tcaccacgga
     1261 atggcatcaa tgacaggagg acagcaaatg ggtcgtgatt tgtatgacga cgatgataaa
     1321 gatttggcta ctatggttga ttcaagccgc cgtaaatgga ataagactgg tcatgctgta
     1381 cgagctattg gaaggttaag ttcccttgaa aatgtttata taaaagcaga taagcagaaa
     1441 aatggtatca aagctaattt tcatatacga cataacattg aagatggtgg tgttcaactt
     1501 gcatatcact atcaacagaa tactcctatt ggcgacggac ctgttttact tcctgataat
     1561 cattatctat ctgtccaatc aaaactgtct aaagacccaa acgaaaaacg tgatcatatg
     1621 gttcttttgg aatttgtgac ggctgccggt attacattag ggatggatga gctttacaaa
     1681 gggggtaccg gtggtagtat ggtttcgaaa ggagaagagt tgtttactgg agtagtacca
     1741 attctggttg aattagacgg agatgttaat ggtcataaat tttcagtttc tggagaaggg
     1801 gaaggagatg ctacatatgg aaaacttacg cttaagttca tatgcaccac aggaaaattg
     1861 cctgttccat ggcccacctt ggttacaaca ttaacatatg gtgtccaatg ttttagtaga
     1921 tatccagatc atatgaaaca acatgacttt ttcaaatctg caatgccaga aggctatatt
     1981 caagagagaa ctatattttt taaggatgat ggaaactata agacaagagc tgaagtgaaa
     2041 tttgaaggag atactttagt gaatcgcata gaacttaaag gtatagactt taaagaggat
     2101 ggcaatattt taggtcacaa attagagtac aatctacctg atcaactaac agaggaacag
     2161 attgcagagt tcaaagaggc attttcactt tttgataaag atggcgatgg aacaatcact
     2221 actaaggaat taggtactgt aatgagatca cttggtcaaa atcctactga agcggaatta
     2281 caagatatga ttaacgaagt agatgcagat ggggatggaa ctatagactt tccggaattt
     2341 ttaacaatga tggcacgaaa aatgaaatac cgtgatactg aagaggaaat tagagaagca
     2401 tttggtgtat ttgacaaaga tggaaacggt tacatttctg ctgctgaact aagacatgtt
     2461 atgacaaact taggagaaaa actgactgat gaagaggtcg acgaaatgat aagagaagcc
     2521 gatattgatg gagatggtca agttaattac gaagagtttg ttcaaatgat gacagcaaag
     2581 taagaattca caattcgatt atatttatac tggactattt ttacatctgt tcggttattt
     2641 tcacatttat ttttctatat atatcttata aacgttttaa aacccatgta atttttgtta
     2701 agctgtaata taaaagacgt cctaacaaac ttcttttatt actgaatttc ctttaattat
     2761 aataaataac aagttttaaa ataaattcag gcaattaagg cgctcctgag gtactaaaat
     2821 taatgtaaac atttaaaatt aacttggatg gtcttaagta ctgtactcgt gattttgtta
     2881 tactttatta ttagaaaagt cgtctattaa ctttttgttc cttaatttac ttgattaaat
     2941 tgtcgcttaa tttatcaaat caggttttgc gcgttatttt agagaaaaac ttattagaaa
     3001 aatgaataag caaagtttag gctaacatgt ttttttatta ttttaaatag ttcaagtcaa
     3061 tgacgtataa accagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3121 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3181 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3241 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3301 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3361 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3421 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3481 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3541 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3601 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3661 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3721 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3781 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3841 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3901 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3961 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4021 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4081 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4141 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4201 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4261 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4321 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4381 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4441 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4501 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4561 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4621 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4681 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4741 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4801 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4861 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4921 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4981 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5041 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5101 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5161 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5221 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5281 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5701 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaac
//
