LOCUS       Exported                6404 bp ds-DNA     circular SYN 16-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6404)
  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..6404
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     29..52
                     /label=f.ITR_seq.01.30
     protein_bind    103..124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        139..169
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    177..193
                     /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)."
     primer_bind     201..217
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     201..217
                     /label=M13R
     misc_feature    255..422
                     /label=ITR
                     /note="ITR"
     misc_feature    451..469
                     /label=AA0208  SacI 480 f
                     /note="AA0208  SacI 480 f"
     misc_feature    487..974
                     /label=synapsin promoter
                     /note="synapsin promoter"
     misc_feature    487..507
                     /label=Syn speI 5'f
                     /note="Syn speI 5'f"
     misc_feature    505..523
                     /label=syn forward primer cactggacaagcacccaac
                     /note="syn forward primer cactggacaagcacccaac"
     primer_bind     763..783
                     /label=f.hsyn.01.16
     primer_bind     complement(976..1001)
                     /label=r.pCAG.01.1
     intron          977..1109
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    complement(1095..1114)
                     /label=intron AscI 3'r
                     /note="intron AscI 3'r"
     protein_bind    complement(1122..1155)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    1216
                     /label=C to A switch indicated in sequencing
                     /note="C to A switch indicated in sequencing"
     protein_bind    complement(1218..1251)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     1235..1298
                     /label=yCaMPs-in-pAAV.r
     CDS             complement(1271..2531)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium 
                     sensor"
                     /label=GCaMP7b
                     /translation="DIATMVDSSRRKWNKTGHAVRAIGRLSSLENVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSYQPVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGD
                     ATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQE
                     RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNPPDQLTEEQI
                     AEFKELFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGDGTIDFPEF
                     LTMYARKMKYRDTEEEIREAFGVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIRE
                     ADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1352..1371
                     /label=f.CMPRI.FLEX.seq.2.01.18
     primer_bind     complement(1625..1642)
                     /label=f.Seq-spGCamP-cen.01.37
     misc_feature    1872..1874
                     /label=I250V (I93V, unknown effect, V in all YFPs&sfGFP)
     misc_feature    1935..1937
                     /label=S229A (S72A, yellow mutation, A in all YFPs)
     misc_feature    1947..1949
                     /label=V225L (V68L, from Citrine seq, unknown effect)
     misc_feature    1956..1958
                     /label=T222G (T65G, yellow Chromophore mutation)
     misc_feature    2013..2015
                     /label=F203L (F36L, maturation mutation in Venus)
     primer_bind     2087..2107
                     /label=EGFP-N
     primer_bind     complement(2214..2235)
                     /label=EGFP-C
     primer_bind     complement(2249..2267)
                     /label=f.Seq-GCamP-Cter_f.01.38
     misc_feature    2268..2270
                     /label=K118V (K206V, close to rescue mutation)
     misc_feature    2271..2273
                     /label=S117P (S205P,Kaspar's rexcue muatation)
     misc_feature    2277..2279
                     /label=V115Y (V203Y, pi-stack muation for yel. Chromoph)
     misc_feature    2427..2429
                     /label=K65T (K153T, sf-mutation, actually M in Citrine)
     primer_bind     complement(2503..2550)
                     /label=yCaMP-in-pAAV.f
     regulatory      2516..2525
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    2550..2583
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    2635..2640
                     /label=Former BglII site
                     /note="Former BglII site"
     protein_bind    2654..2687
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2722..2727
                     /label=Former EcoRI site
                     /note="Former EcoRI site"
     misc_feature    2727..2732
                     /label=Former PstI site
                     /note="Former PstI site"
     misc_feature    2759..3347
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     2759..2786
                     /label=WPRE into pUCmm-iCap
     primer_bind     complement(2812..2832)
                     /label=r.WPRE.01.29
     primer_bind     complement(2813..2835)
                     /label=r.WPRE_seq.01.32
     misc_feature    2830..2849
                     /label=WPRE rev WPRE rev ttaaagcagcgtatccacat
                     /note="WPRE rev WPRE rev ttaaagcagcgtatccacat"
     CDS             complement(3230..3241)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3329..3347)
                     /label=r.WPRE into pUCmm-iCap.10.
     polyA_signal    3425..3546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3440..3466)
                     /label=r.SV40poly(A)long.01.12
     primer_bind     complement(3451..3467)
                     /label=r.SV40poly(A).01.6
     primer_bind     3490..3509
                     /label=SV40pA-R
     misc_feature    3670..3837
                     /label=ITR
                     /note="ITR"
     primer_bind     complement(3856..3873)
                     /label=M13F(-21)
     primer_bind     complement(3856..3872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3876..3899)
                     /label=M13F(-47)
     primer_bind     complement(3876..3893)
                     /label=M13F(-41)
     rep_origin      4014..4469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4495..4599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4600..5460
                     /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      5631..6219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
       61 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      121 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      181 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca gatttaatta
      241 aggccttaat taggctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
      301 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
      361 ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta
      421 cgtagccatg ctctaggaag atccgagctc cagtgtgctg gaattcgccc ttttcagtat
      481 ttaaattgac gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg
      541 cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc
      601 cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc
      661 tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc
      721 accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga
      781 taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt
      841 ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagctgtgc tcctgggcac
      901 cgcgcagtcc gcccccgcgg ctcctggcca gaccacccct aggaccccct gccccaagtc
      961 gcagccgttg gatcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     1021 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     1081 tgacatccac tttgcctttc tctccacagg tgatgatccc cataacttcg tataaagtat
     1141 cctatacgaa gttatatcaa aataggaaga ccaatgcttc accatcgacc cgaattgcca
     1201 agcatcacca tcgacacata acttcgtata atgtatgcta tacgaagtta tactagtgct
     1261 agcggatcct acttcgctgt catcatttgt acaaactctt cgtagtttac ctgaccatcc
     1321 ccatcgatgt ctgcttccct gatcatttca tcaacctctt catctgttaa cttctctcca
     1381 aggtttgtca tcacgtggcg aagctctgct gcactgatgt agccattgcc atccttatca
     1441 aacacaccga acgcttctct aatttcttct tccgtgtccc tgtatttcat ttttcttgcg
     1501 tacattgtca ggaactcagg gaagtcgatt gtgccgtcac cgtcggcatc tacttcattg
     1561 atcatgtcct gcagctctgc ttctgtgggg ttctgcccca gagaccgcat caccgtcccc
     1621 agctccttgg ttgttattgt cccatccccg tccttgtcaa atagggagaa aagctcttta
     1681 aattctgcga tctgctcttc agtcagttgg tcaggtgggt tgtactccag cttgtgcccc
     1741 aggatgttgc cgtcctcctt gaagtcgatg cccttaagct cgatgcggtt caccagggtg
     1801 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1861 gtgcgctcct gcacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     1921 tggtcggggt agcgggcgaa gcactgcagg ccgtagccca gggtggtcac gagggtgggc
     1981 cagggcacgg gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg
     2041 gcatcgccct caccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     2101 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catgctccct
     2161 ccggtaccgc ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
     2221 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tcgaaagcac gggctggtag
     2281 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     2341 tagtggtagg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttcgcc
     2401 ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tctcgagtga gctcagccga
     2461 cctatagctc tgactgcgtg acctgtctta ttccacttac gacgtgatga gtcgaccatg
     2521 gtggcgatat caccggtggt accactagta taacttcgta taggatactt tatacgaagt
     2581 tatcattggg attcttccta ttttgatcca agcatcacca tcgaccctct agtccaaatc
     2641 tcaccatcga cccataactt cgtatagcat acattatacg aagttatgtc cctcgaagag
     2701 gttcgaaaca ctgaaaaggg ccaattctgc aaatgatcca ctcgagaagc ttatcgataa
     2761 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2821 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2881 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2941 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3001 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3061 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3121 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3181 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3241 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3301 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     3361 acccgggcgg ccgctcgagc atgcatctag agtcgacccg ggcggccgct tcgagcagac
     3421 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3481 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3541 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     3601 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga
     3661 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     3721 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3781 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagcct
     3841 taattaacct aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3901 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3961 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     4021 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4081 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4141 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4201 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4261 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4321 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4381 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4441 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4561 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4621 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4681 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4741 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4801 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4861 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4921 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4981 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5041 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5101 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5161 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5221 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5281 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5341 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5401 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5461 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5581 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5761 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6241 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6301 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6361 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcg
//
