LOCUS       Exported                7130 bp DNA     circular SYN 17-MAR-2022
DEFINITION  pAAV-MCS from stratagen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  AUTHORS   Demo User
  TITLE     Direct Submission
  JOURNAL   Exported Mar 17, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
COMMENT     Createdd by Wei.  
            
            Storage Type: Basic
            Original Author: 
            UNKNOWN
            mCherry were PCR cloned as NheI-EcoRI-ATG-mCherry-TAG-HindIII from 
            pACK-GCaMP2.0-NLS-mCherry from Eckehard Freitag248|mCherry cds were 
            obtained from MPI-Munich.248|http://www.informaxinc.com/
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     promoter        158..1446
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    1161..1178
                     /label=ACK-F2 Primer
                     /note="ACK-F2 primer binding site"
                     /note="This feature has 2 segments:
                        1: 1161 .. 1162 / #31849b
                        2: 1163 .. 1178 / #31849b"
     regulatory      1457..1466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             1459..2508
                     /codon_start=1
                     /label=MCU
                     /note="color: #708090"
                     /translation="MAAAAGRSLLLLLCSRGGGGGAGGCGALTAGCFPGLGVSRHRPHQ
                     QHRTAHQRPASWQSVGAAYCSTVVPSDDVTVVYQNGLPVISVRLPSRRERCQFTLKPIS
                     DSVGVFLRQLQEEDRGIDRVAIYSPDGVRVAASTGIDLLLLDDFKLVINDLTYHVRPPK
                     RDLLSHEDAATLNDVKTLVQQLYTTLCIEQHQLNKERELVERLEDLKQQLAPLEKVRIE
                     ISRKAEKRTTLVLWGGLAYMATQFGILARLTWWEYSWDIMEPVTYFITYGSAMAMYAYF
                     VMTRQEYVYPEARDRQYLLFFHKGAKKSRFDLEKYNQLKDAIAQAEMDLKRLRDPLQVH
                     LPLRQIGEKE"
     CDS             2515..3234
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 2515 .. 2517 / #05fd14
                        2: 2518 .. 2520 / #05fd14 / 1a
                        3: 2521 .. 3234 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2516..2518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     misc_feature    3259..3847
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     3345..3363
                     /label=CAAV-R
                     /note="CAAV-R primer"
                     /note="color: #a020f0; direction: RIGHT"
     CDS             complement(3730..3741)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 3729"
                     /translation="IEGR"
     primer_bind     complement(3850..3866)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     polyA_signal    3877..4353
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   4393..4533
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      4608..5063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     TATA_signal     5277..5283
                     /note="color: #a6acb3"
     promoter        5345..5449
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5450..6310
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5450 .. 5518 / #ccffcc / signal sequence
                        2: 5519 .. 6310 / #ccffcc
                     Cleavage site after base 5518"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6481..7069
                     /direction=RIGHT
                     /label=ori
                     /note="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 cgcgtaacat tatggcctta ggtcacttca
      181 tctccatggg gttcttcttc tgattttcta gaaaatgaga tgggggtgca gagagcttcc
      241 tcagtgacct gcccagggtc acatcagaaa tgtcagagct agaacttgaa ctcagattac
      301 taatcttaaa ttccatgcct tgggggcatg caagtacgat atacagaagg agtgaactca
      361 ttagggcaga tgaccaatga gtttaggaaa gaagagtcca gggcagggta catctacacc
      421 acccgcccag ccctgggtga gtccagccac gttcacctca ttatagttgc ctctctccag
      481 tcctaccttg acgggaagca caagcagaaa ctgggacagg agccccagga gaccaaatct
      541 tcatggtccc tctgggagga tgggtgggga gagctgtggc agaggcctca ggaggggccc
      601 tgctgctcag tggtgacaga taggggtgag aaagcagaca gagtcattcc gtcagcattc
      661 tgggtctgtt tggtacttct tctcacgcta aggtggcggt gtgatatgca caatggctaa
      721 aaagcaggga gagctggaaa gaaacaagga cagagacaga ggccaagtca accagaccaa
      781 ttcccagagg aagcaaagaa accattacag agactacaag ggggaaggga aggagagatg
      841 aattagcttc ccctgtaaac cttagaaccc agctgttgcc agggcaacgg ggcaatacct
      901 gtctcttcag aggagatgaa gttgccaggg taactacatc ctgtctttct caaggaccat
      961 cccagaatgt ggcacccact agccgttacc atagcaactg cctctttgcc ccacttaatc
     1021 ccatcccgtc tgttaaaagg gccctatagt tggaggtggg ggaggtagga agagcgatga
     1081 tcacttgtgg actaagtttg ttcgcatccc cttctccaac cccctcagta catcaccctg
     1141 ggggaacagg gtccacttgc tcctgggccc acacagtcct gcagtattgt gtatataagg
     1201 ccagggcaaa gaggagcagg ttttaaagtg aaaggcaggc aggtgttggg gaggcagtta
     1261 ccggggcaac gggaacaggg cgtttcggag gtggttgcca tggggacctg gatgctgacg
     1321 aaggctcgcg aggctgtgag cagccacagt gccctgctca gaagccccaa gctcgtcagt
     1381 caagccggtt ctccgtttgc actcaggagc acgggcaggc gagtggcccc tagttctggg
     1441 ggcagcgggg gatccaccat ggcggccgcc gcaggtagat cgctcctgct gctgctctgc
     1501 tcccggggcg gcggcggggg cgccggcggc tgcggagcgc tgactgccgg ctgctttccc
     1561 gggctgggcg tcagccgcca ccggccgcac cagcagcacc ggacggcaca ccagaggccg
     1621 gcttcctggc agagcgtggg agccgcatat tgcagtacgg ttgtgccctc tgatgacgtg
     1681 acggtggttt accaaaatgg attaccggtg atatctgtga ggctaccctc tcgacgcgag
     1741 cgctgccaat tcacactcaa gcctatctct gactcagtcg gcgtgttttt acgacaactg
     1801 caagaggagg atcggggaat cgaccgggtc gccatctact caccagatgg cgttcgagtc
     1861 gctgcctcca cggggataga cctcctgctc ctcgatgact ttaagcttgt cattaatgac
     1921 ttaacatacc acgtacggcc accaaagaga gacctcctaa gccatgaaga tgcagcgacg
     1981 ctgaacgacg tgaagaccct ggtccagcag ctgtacacca cactgtgcat tgagcagcat
     2041 cagcttaaca aagagcggga gctcgtggag aggttagagg acctcaagca gcagctggcc
     2101 cccctggaga aggtacgaat tgaaattagc agaaaagcag agaagaggac cactctggtg
     2161 ctgtggggag gcctggccta catggccacc cagtttggca ttctggcccg gctcacctgg
     2221 tgggagtact cgtgggacat catggagccc gtcacctact tcatcacgta cggaagcgcc
     2281 atggccatgt atgcgtattt tgtaatgacg cgccaggaat atgtttatcc agaagccaga
     2341 gacagacaat acttattatt tttccataaa ggagccaaaa agtcacgttt cgacctagag
     2401 aaatacaatc agctcaagga tgcaattgct caggcagaaa tggatcttaa gagactgaga
     2461 gacccattac aagtacacct gcccctccga cagatcggag aaaaggaagc tagcatggtg
     2521 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2581 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2641 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2701 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2761 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2821 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2881 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2941 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3001 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3061 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3121 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3181 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagaattc
     3241 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3301 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3361 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3421 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3481 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3541 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3601 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3661 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3721 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3781 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3841 tccccgcatc gataccgtcg acctcgagag atctacgggt ggcatccctg tgacccctcc
     3901 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     3961 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     4021 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     4081 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     4141 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     4201 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4261 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     4321 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     4381 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4441 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4501 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4561 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4621 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4681 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4741 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4801 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4861 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4921 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     4981 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5041 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5101 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5161 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5221 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5281 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5461 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5521 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5581 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5641 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5701 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5761 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5821 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5881 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5941 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6001 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6061 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6121 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6181 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6241 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6301 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6361 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6421 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6481 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6541 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6601 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6661 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6721 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6781 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6841 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6901 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6961 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7021 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7081 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
