LOCUS       Exported                6973 bp DNA     circular SYN 30-APR-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6973)
  AUTHORS   Frank Voss
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 14, 2021 from SnapGene Viewer 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6973
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="AAV2 ITR
                     inverted terminal repeat of adeno-associated virus serotype
                     2"
                     /note="color: #a6acb3"
     promoter        242..1419
                     /note="EF-1-alpha promoter
                     strong constitutive promoter for human elongation
                     factor EF-1-alpha"
                     /note="color: #ffffff"
     intron          473..1410
                     /note="EF-1-alpha intron A
                     intron upstream of the start codon of human
                     EF-1-alpha"
                     /note="color: #------"
     protein_bind    1426..1459
                     /label=FLP recombinase from the Saccharomyces
                     cerevisi
                     /bound_moiety="FLP recombinase from the 
                     Saccharomycescerevisiae 2u plasmid"
                     /note="F14"
                     /note="color: #31849b"
     protein_bind    1510..1543
                     /label=FLP recombinase from the Saccharomyces
                     cerevisi
                     /bound_moiety="FLP recombinase from the 
                     Saccharomycescerevisiae 2u plasmid"
                     /note="FRT (minimal)
                     supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
                     /note="color: #31849b"
     CDS             complement(1557..2909)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=jGCaMP7b
                     /label=GCaMP6f
                     /note="color: #993366"
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNPPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMYARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     2116..2135
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     2375..2396
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(2502..2523)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: #a020f0; direction: LEFT"
     regulatory      2804..2813
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #ffffff"
     regulatory      2906..2915
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #ffffff"
     protein_bind    complement(2956..2989)
                     /label=FLP recombinase from the Saccharomyces
                     cerevisi
                     /bound_moiety="FLP recombinase from the 
                     Saccharomycescerevisiae 2u plasmid"
                     /note="F14"
                     /note="color: #31849b; direction: LEFT"
     protein_bind    complement(3040..3073)
                     /label=FLP recombinase from the Saccharomyces
                     cerevisi
                     /bound_moiety="FLP recombinase from the 
                     Saccharomycescerevisiae 2u plasmid"
                     /note="FRT (minimal)
                     supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
                     /note="color: #31849b; direction: LEFT"
     misc_feature    3100..3688
                     /label=WPRE
                     /note="WPRE
                     woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /note="color: #a6acb3"
     CDS             complement(3571..3582)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa recognition and cleavage site
                     /note="Factor Xa site"
                     /note="color: #993366"
                     /translation="IEGR"
     polyA_signal    3720..4196
                     /label=hGH poly(A) signal
                     /note="hGH poly(A) signal
                     human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   4236..4376
                     /label=AAV2 ITR
                     /note="AAV2 ITR
                     inverted terminal repeat of adeno-associated virus serotype
                     2"
                     /note="color: #a6acb3"
     rep_origin      4451..4906
                     /label=f1 ori
                     /note="f1 ori
                     f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5188..5292
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
                     /note="color: #ffffff"
     CDS             5293..6153
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=Ampicilin
                     /note="AmpR
                     confers resistance to ampicillin, carbenicillin, and 
                     related antibiotics"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6324..6912
                     /label=ori
                     /note="ori
                     high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccgaagt tcctattcta
     1441 tcagaagtat aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga
     1501 aacctcttag aagttcctat tctctagaaa gtataggaac ttcgaattct tacttttcac
     1561 ttcgctgtca tcatttgtac aaactcttcg tagtttacct gaccatcccc atcgatgtct
     1621 gcttccctga tcatttcatc aacctcttca tctgttaact tctctccaag gtttgtcatc
     1681 acgtggcgaa gctctgctgc actgatgtag ccattgccat ccttatcaaa cacaccgaac
     1741 gcttctctaa tttcttcttc cgtgtccctg tatttcattt ttcttgcgta cattgtcagg
     1801 aactcaggga agtcgattgt gccgtcaccg tcggcatcta cttcattgat catgtcctgc
     1861 agctctgctt ctgtggggtt ctgccccaga gaccgcatca ccgtccccag ctccttggtt
     1921 gttattgtcc catccccgtc cttgtcaaat agggagaaaa gctctttaaa ttctgcgatc
     1981 tgctcttcag tcagttggtc aggtgggttg tactccagct tgtgccccag gatgttgccg
     2041 tcctccttga agtcgatgcc cttaagctcg atgcggttca ccagggtgtc gccctcgaac
     2101 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     2161 atgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     2221 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
     2281 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctca
     2341 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     2401 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tgctccctcc ggtaccgccc
     2461 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     2521 atgtgatcgc gcttctcgtt ggggtctttc gaaagtttgg actgcacgct caggtagtgg
     2581 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtaggcg
     2641 agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttcgcctt gatgccgttc
     2701 ttctgcttgt cggccttgat atagacgttc tcgagtgagc tcagccgacc tatagctctg
     2761 actgcgtgac ctgtcttatt ccacttacga cgtgatgagt cgaccatggt ggcgagatcc
     2821 ttatcgtcat cgtcgtacag atcccgaccc atttgctgtc caccagtcat gctagccata
     2881 ccatgatgat gatgatgatg agaacccatg gtggcgggtg gatcccggct agctacgtag
     2941 gcgcgcctta attaagaagt tcctatactt ctgatagaat aggaacttct tgccttaacc
     3001 cagaaattat cactgttatt ctttagaatg gtgcaaagag aagttcctat actttctaga
     3061 gaataggaac ttccagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     3121 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3181 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3241 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3301 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3361 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3421 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3481 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     3541 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3601 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3661 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg
     3721 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     3781 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     3841 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     3901 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     3961 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     4021 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     4081 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     4141 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     4201 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4261 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4321 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4381 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4441 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4501 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4561 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4621 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4681 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4741 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     4801 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4861 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4921 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4981 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5041 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5101 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5161 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5221 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5281 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5341 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5401 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5461 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5521 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5581 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5641 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5701 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5761 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5821 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5881 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5941 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6001 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6061 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6121 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6181 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6241 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6301 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6361 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6421 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6481 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6541 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6601 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6661 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6721 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6781 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6841 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6901 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6961 tttgctcaca tgt
//
