LOCUS       Exported                5663 bp ds-DNA     circular SYN 30-OCT-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5663)
  AUTHORS   Manuel Mohr
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Oct 30, 2019 from SnapGene 5.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5663
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    56..222
                     /label=ITR
                     /note="ITR"
     misc_feature    239..244
                     /label=BglII
                     /note="BglII"
     promoter        266..713
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             770..2019
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium 
                     sensor"
                     /label=GCaMP7b
                     /translation="MVDSSRRKWNKTGHAVRAIGRLSSLENVYITADKQKNGIKANFKI
                     RHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSYQPVLSKDPNEKRDHMVLLEFVTA
                     AGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYG
                     KLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIF
                     FKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNPPDQLTEEQIAEFK
                     ELFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGDGTIDFPEFLTMY
                     ARKMKYRDTEEEIREAFGVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADID
                     GDGQVNYEEFVQMMTAK"
     regulatory      770..774
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    861..863
                     /label=K65T (K153T, sf-mutation, actually M in Citrine)
     misc_feature    1011..1013
                     /label=V115Y (V203Y, pi-stack muation for yel. Chromoph)
     misc_feature    1017..1019
                     /label=S117P (S205P,Kaspar's rexcue muatation)
     misc_feature    1020..1022
                     /label=K118V (K206V, close to rescue mutation)
     misc_feature    1275..1277
                     /label=F203L (F36L, maturation mutation in Venus)
     misc_feature    1332..1334
                     /label=T222G (T65G, yellow Chromophore mutation)
     misc_feature    1341..1343
                     /label=V225L (V68L, from Citrine seq, unknown effect)
     misc_feature    1353..1355
                     /label=S229A (S72A, yellow mutation, A in all YFPs)
     misc_feature    1416..1418
                     /label=I250V (I93V, unknown effect, V in all YFPs&sfGFP)
     misc_feature    2050..2638
                     /label=WPRE
                     /note="WPRE"
     misc_feature    2186..2206
                     /label=Mao 115r
                     /note="Mao 115r"
     CDS             complement(2521..2532)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2641..2657)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2687..2808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2932..3098
                     /label=ITR
                     /note="ITR"
     primer_bind     complement(3118..3134)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3276..3731
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3757..3861
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3862..4722
                     /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"
     rep_origin      4893..5481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc
       61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
      121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
      241 atctctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg
      301 gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca acccccattc
      361 cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt
      421 gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc
      481 accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc
      541 ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga
      601 ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc
      661 tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt
      721 caagctgcta gcaaggatcc gaattcacta gtctcgagga tatcgccacc atggtcgact
      781 catcacgtcg taagtggaat aagacaggtc acgcagtcag agctataggt cggctgagct
      841 cactcgagaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gcgaacttca
      901 agatccgcca caacatcgag gacggcggcg tgcagctcgc ctaccactac cagcagaaca
      961 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagcccg
     1021 tgctttcgaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1081 ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatgg
     1141 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1201 acgtaaacgg ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc acctacggca
     1261 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1321 tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1381 acgacttctt caagtccgcc atgcccgaag gctacgtgca ggagcgcacc atcttcttca
     1441 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1501 accgcatcga gcttaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1561 tggagtacaa cccacctgac caactgactg aagagcagat cgcagaattt aaagagcttt
     1621 tctccctatt tgacaaggac ggggatggga caataacaac caaggagctg gggacggtga
     1681 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag
     1741 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgtac gcaagaaaaa
     1801 tgaaatacag ggacacggaa gaagaaatta gagaagcgtt cggtgtgttt gataaggatg
     1861 gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt
     1921 taacagatga agaggttgat gaaatgatca gggaagcaga catcgatggg gatggtcagg
     1981 taaactacga agagtttgta caaatgatga cagcgaagta aggtgtacaa agatatcaag
     2041 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2101 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2161 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2221 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2281 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2341 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2401 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2461 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2521 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2581 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     2641 cgataccgtc gacctcgacc cgggcggccg cttcgagcag acatgataag atacattgat
     2701 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2761 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2821 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     2881 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     2941 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     3001 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3061 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact
     3121 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3181 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3241 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     3301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3421 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3481 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3541 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3601 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3661 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3721 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3781 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3841 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3901 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3961 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4021 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4081 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4141 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4201 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4261 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4321 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4381 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4441 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4501 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4561 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4621 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4681 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4741 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4801 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4861 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4921 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4981 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5041 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5101 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5161 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5221 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5281 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5341 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5401 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5461 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5521 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5581 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5641 tcagtgagcg aggaagcgga aga
//
