LOCUS       Exported                7736 bp DNA     circular SYN 25-MAR-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7736)
  AUTHORS   Anna M. H. Hertle
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7736
                     /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"
     primer_bind     complement(229..247)
                     /label=U6_rev
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     1301..1322
                     /label=CKIIa_for
                     /note="color: #a020f0; direction: RIGHT"
     CDS             1489..3840
                     /codon_start=1
                     /label=4mt.D3cpv
                     /note="color: #993366"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDPMSVLTPLLLRGLT
                     GSARRLPVPRAKIHSLGKLMSVLTPLLLRGLTGSARRLPVPRAKIHSLGDPMSVLTPLL
                     LRGLTGSARRLPVPRAKIHSLGKLATMVSKGEELFTGVVPILVELDGDVNGHRFSVSGE
                     GEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEG
                     YVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNV
                     YITADKQKNGIKAHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSK
                     DPNEKRDHMVLLEFVTAARMHDQLTEEQIAEFKEAFSLLDKDGDGTITTKELGTALRSL
                     GQNPTEAELQDMINEVDADGNGTIYFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNG
                     YISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAKGGKRRWQKT
                     GHAVRAFGRLKKISSSGALELMDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSK
                     DPNEKRDHMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNG
                     HKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDF
                     FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE
                     YNYNSHNVYITADKQKNGIKANFKIRHNIE"
     CDS             1489..1563
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /note="color: #993366
                     Cleavage site after base 1563"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1585..1659
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /note="color: #993366
                     Cleavage site after base 1659"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1681..1755
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /note="color: #993366
                     Cleavage site after base 1755"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1777..1851
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /note="color: #993366
                     Cleavage site after base 1851"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      1873..1882
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             1879..2565
                     /codon_start=1
                     /label=ECFP
                     /note="coding sequence for cyan fluorescent protein"
                     /note="color: #00ccff"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAR"
     CDS             3103..3837
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=cpVenus
                     /note="circularly permuted venus"
                     /note="This feature has 2 segments:
                        1: 3103 .. 3318 / #99cc00
                        2: 3319 .. 3837 / #99cc00 / VN173"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     CDS             3319..3837
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /note="This feature has 3 segments:
                        1: 3319 .. 3321 / #e1ff2f
                        2: 3322 .. 3324 / #e1ff2f / 1a
                        3: 3325 .. 3837 / #e1ff2f"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     misc_feature    3865..4453
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(3951..3969)
                     /label=WPRE_rev
                     /note="color: #a020f0; direction: LEFT"
     CDS             complement(4336..4347)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4335"
                     /translation="IEGR"
     primer_bind     complement(4456..4472)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     polyA_signal    4483..4959
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   4999..5139
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      5214..5669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5951..6055
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             6056..6916
                     /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: 6056 .. 6124 / #ccffcc / signal sequence
                        2: 6125 .. 6916 / #ccffcc
                     Cleavage site after base 6124"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7087..7675
                     /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 ttcacatccc cttctccaac cccctcagta catcaccctg
     1141 ggagaacaag gtccacttgc ttctgggccc acacagtcct gcagtattgt gtatataagg
     1201 ccagggcaac ggaggagcag gttttaagtg 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 gatccaccgg tcgactcact atagggagac ccaagcttat gtccgtcctg
     1501 acgccgctgc tgctgcgggg cttgacaggc tcggcccggc ggctcccagt gccgcgcgcc
     1561 aagatccatt cgttggggga tcccatgtcc gtcctgacgc cgctgctgct gcggggcttg
     1621 acaggctcgg cccggcggct cccagtgccg cgcgccaaga tccattcgtt ggggaagctt
     1681 atgtccgtcc tgacgccgct gctgctgcgg ggcttgacag gctcggcccg gcggctccca
     1741 gtgccgcgcg ccaagatcca ttcgttgggg gatcccatgt ccgtcctgac gccgctgctg
     1801 ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc cgcgcgccaa gatccattcg
     1861 ttggggaagc ttgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1921 atcctggtcg agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc
     1981 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2041 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc
     2101 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2161 caggagcgta ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2221 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2281 ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc
     2341 gccgacaagc agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac
     2401 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2461 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2521 aagcgcgatc acatggtcct gctggagttc gtgaccgccg cccgcatgca tgaccaactg
     2581 acagaagagc agattgcaga gttcaaagaa gccttctcat tattggacaa ggatggggac
     2641 ggcaccatca ccacaaagga acttggcacc gctctgaggt cgcttggaca aaacccaacg
     2701 gaagcagaat tgcaggatat gatcaatgaa gtcgatgctg atggcaatgg aacgatttac
     2761 tttcctgaat ttcttactat gatggctaga aaaatgaagg acacagacag cgaagaggaa
     2821 atccgagaag cattccgtgt ttttgacaag gatgggaacg gctacatcag cgctgctgaa
     2881 ttacgtcacg tcatgacaaa cctcggggag aagttaacag atgaagaagt tgatgaaatg
     2941 ataagggaag cagatatcga tggtgatggc caagtaaact atgaagagtt tgtacaaatg
     3001 atgacagcaa agggggggaa gaggcgctgg cagaaaacag gccatgctgt ccgagcattc
     3061 ggaagactga agaagatctc cagctccggg gcactggagc tcatggacgg cggcgtgcag
     3121 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3181 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3241 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3301 aagggtggca gcggtggcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3361 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3421 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
     3481 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc
     3541 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3601 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3661 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3721 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
     3781 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgagtaa
     3841 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3901 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3961 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4021 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4081 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4141 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4201 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4261 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4321 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4381 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4441 gccgcctccc cgcatcgata ccgtcgacct cgagagatct acgggtggca tccctgtgac
     4501 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     4561 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4621 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4681 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4741 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4801 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4861 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4921 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4981 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     5041 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     5101 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     5161 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     5221 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5281 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5341 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5401 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     5461 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5521 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     5581 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5641 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5701 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5761 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5821 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5881 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5941 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6001 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6061 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6121 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6181 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6241 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6301 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6361 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6421 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6481 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6541 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6601 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6661 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6721 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6781 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6841 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6901 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6961 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7201 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     7261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7381 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7441 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7501 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7561 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7621 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7681 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
