LOCUS       40924_22453        5308 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of ultrasensitive protein calcium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5308)
  AUTHORS   Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, 
            Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, 
            Kim DS
  TITLE     Ultrasensitive fluorescent proteins for imaging neuronal activity.
  JOURNAL   Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
  PUBMED    23868258
REFERENCE   2  (bases 1 to 5308)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5308)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2013-07-18- 18"; volume: "499"; issue: "7458"; 
            pages: "295-300"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(236..340)
                     /label=AmpR promoter
     rep_origin      367..822
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    complement(829..950)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1075..2424)
                     /codon_start=1
                     /label=GCaMP6s
                     /note="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     promoter        complement(2478..2681)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(2682..2985)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3160..3748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(4077..4124)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4359..5150)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(join(5185..5308,1..234))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      181 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
      241 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      301 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      361 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcctta agatacattg
      841 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      901 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      961 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1021 aaaacctcta caaatgtggt atggctgatt atgatctaga gtcgcggccg ctcacttcgc
     1081 tgtcatcatt tgtacaaact cttcgtagtt tacctgacca tccccatcga tgtctgcttc
     1141 cctgatcatt tcatcaacct cttcatctgt taacttctct ccaaggtttg tcatcacgtg
     1201 gcgaagctct gctgcactga tgtagccatt gccatcctta tcaaacacac cgaacgcttc
     1261 tctaatttct tcttccgtgt ccctgtattt catttttctt gccatcattg tcaggaactc
     1321 agggaagtcg attgtgccgt caccgtcggc atctacttca ttgatcatgt cctgcagctc
     1381 tgcttctgtg gggttctgcc ccagagaccg catcaccgtc cccagctcct tggttgttat
     1441 tgtcccatcc ccgtccttgt caaataggga gaaagcctct ttaaattctg cgatctgctc
     1501 ttcagtcagt tggtccggca ggttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     1561 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     1621 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggatgta
     1681 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     1741 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     1801 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcaccctc
     1861 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     1921 caccccggtg aacagctcct cgcccttgct caccatgctc cctccggtac cgcccttgta
     1981 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     2041 atcgcgcttc tcgttggggt ctttcgaaag tttggactgc acgctcaggt agtggttgtc
     2101 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt aggcgagctg
     2161 cacgccgccg tcctcgatgt tgtggcggat gtggaagttc gccttgatgc cgttcttctg
     2221 cttgtcggcc ttgatataga cgttctcgag tgagctcagc cgacctatag ctctgactgc
     2281 gtgacctgtc ttattccact tacgacgtga tgagtcgacc atggtggcga gatccttatc
     2341 gtcatcgtcg tacagatccc gacccatttg ctgtccacca gtcatgctag ccataccatg
     2401 atgatgatga tgatgagaac ccatggtggc gagatctgag tccggtagcg ctagcggatc
     2461 tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc
     2521 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     2581 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     2641 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     2701 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     2761 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     2821 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     2881 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     2941 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     3001 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat
     3061 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3601 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3901 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     3961 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     4021 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     4081 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     4141 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     4201 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     4261 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     4321 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     4381 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     4441 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     4501 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     4561 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     4621 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     4681 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     4741 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     4801 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     4861 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     4921 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     4981 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     5041 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     5101 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     5161 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     5221 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     5281 tagtcagcca tggggcggag aatgggcg
//
