LOCUS Exported 5663 bp ds-DNA circular SYN 30-OCT-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct 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="synthetic DNA construct" /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=jGCaMP7f /translation="MVDSSRRKWNKTGHAVRAIGRLSSLENVYITADKQKNGIKANFKI RHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSYQPVLSKDPNEKRDHMVLLEFVTA AGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYG KLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIF FKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFK ELFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGDGTIDFPEFLTMM ARKMKYTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADID GDGQVNYEEFVQMMTAK" regulatory 770..774 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_difference 822..824 /label=difference between s and f misc_feature 861..863 /label=K65T (K153T, sf-mutation, actually M in Citrine) misc_feature 900..902 /label=difference between s and f 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_difference 1572..1574 /label=T302L (GCaMP7f mutation) misc_difference 1575..1577 /label=R303P (GCaMP7f mutation) misc_difference 1617..1618 /label=A317L (GCaMP7f mutation) misc_difference 1806..1808 /label=D380Y (GCaMP7s mutation) misc_difference 1809..1811 /label=difference between s and f misc_difference 1815..1817 /label=difference between s and f misc_difference 1842..1844 /label=difference between s and f 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 cctgccggac caactgactg aagagcagat cgcagaattt aaagagctgt 1621 tctccctatt tgacaaggac ggggatggga caataacaac caaggagctg gggacggtga 1681 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag 1741 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa 1801 tgaaatacac agacagtgaa gaagaaatta gagaagcgtt ccgtgtgttt 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 //