LOCUS       Exported                7016 bp ds-DNA     circular SYN 14-OCT-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7016)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Apr 16, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7016
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     139..170
                     /label=Юра*F_CamkII_Mlu
     promoter        145..1437
                     /label=CaMKII
                     /label=a-CaMKII(1.3)
                     /note="/vntifkey=21"
     primer_bind     431..448
                     /label=Юра*F_CamK
     primer_bind     1074..1099
                     /label=CaMK-sh_F_Xba-Mlu
     primer_bind     1400..1416
                     /label=Лена*pLenti-CamK-F
     primer_bind     complement(1420..1443)
                     /label=Юра*R_CamkII_Nhe
     primer_bind     complement(1420..1438)
                     /label=CamkII_R_Kpn
     primer_bind     1438..1460
                     /label=RFP-F-NheI
     primer_bind     1442..1460
                     /label=mRuby2-NheI-Fw
     primer_bind     1442..1460
                     /label=Катя*mRuby2-NheI-Fw
     primer_bind     1442..1459
                     /label=RFP_F_EcoRI
     regulatory      1442..1451
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1448..2158
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(1803..1830)
                     /label=TagBFP-seq-rev62
     primer_bind     1803..1830
                     /label=TagBFP-seq-dir62
     primer_bind     complement(2135..2164)
                     /label=RFP-R-BamHI
     primer_bind     complement(2135..2158)
                     /label=RFP_R_HindIII
     primer_bind     2159..2176
                     /label=p2A-BamHI-Fw
     primer_bind     2159..2176
                     /label=Катя*p2A-BamHI-Fw
     CDS             2168..2224
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=p2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     2205..2248
                     /label=Юра*F_DAO_overlap
     primer_bind     complement(2205..2224)
                     /label=p2A-Rv
     primer_bind     complement(2205..2224)
                     /label=Катя*p2A-Rv
     CDS             2225..3319
                     /codon_start=1
                     /label=DAAOmut
                     /translation="HSQKRVVVLGSGVIGLSSALILARKGYSVHILARDLPEDVSSQTF
                     ASPWAGANWTPFMTLTDGPRQAKWEESTFKKWVELVPTGHAMWLKGTRRFAQNEDGLLG
                     HWYKDITPNYRPLPSSECPPGAIGVTYDTLSVHAPKYCQYLARELQKLGATFERRTVTS
                     LEQAFDGADLVVNATGLGAKSIAGIDDQAAEPIRGQTVLVKSPCKRCTMDSSDPASPAY
                     IIPRPGGEVICGGTYGVGDWDLSVNPETVQRILKHCLRLDPTISSDGTIEGIEVLRHNV
                     GLAPARRGGPRVEAERIVLPLDRTKSPLSLGRGSARAAKEKEVTLVHAYGFSSAGYQQS
                     WGAAEDVAQLVDEAFQRYHGAARES"
     primer_bind     2240..2257
                     /label=Юльчи и Фены, cat#180
     primer_bind     complement(2757..2773)
                     /label=DAO-MidR, Юльчи и Фены, cat#47
     primer_bind     2757..2773
                     /label=DAO-MidF, Юльчи и Фены, cat#46
     primer_bind     2858..2876
                     /label=DAO-MV-F, Юльчи и Фены, cat#182
     primer_bind     3064..3094
                     /label=Юльчи и Фены, cat#259
     primer_bind     complement(3074..3089)
                     /label=DAAOmut_R
     misc_feature    3074..3075
                     /label=mut
     primer_bind     complement(3259..3275)
                     
     primer_bind     complement(3294..3319)
                     /label=R_DAO_St_EcoRI
     primer_bind     complement(3303..3355)
                     /label=Юльчи и Фены, cat#101
     CDS             3320..3346
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(3332..3349)
                     /label=Юльчи и Фены, cat#92
     misc_feature    3362..3950
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3365..3386)
                     /label=pLenti-R #136
     primer_bind     complement(3412..3431)
                     /label=pLu-Far-R #143
     primer_bind     complement(3415..3435)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(3427..3448)
                     /label=pLVT1-WPRE-Rv (Дина)
     CDS             complement(3833..3844)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4119..4200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4156..4175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4210..4229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4279..4419
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4279..4408
                     /label=AAV2 ITR
     rep_origin      4494..4949
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4581..4600)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4791..4812
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4966..4985)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5085..5107
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5145..5163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5231..5335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5336..6196
                     /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"
     primer_bind     complement(5554..5573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6367..6955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6817..6826)
                     /label=DAO-MV-F, Юльчи и Фены, cat#182
     primer_bind     6856..6875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagcgct
     1441 agccaccatg gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta
     1501 catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc
     1561 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc
     1621 cttcgacatc ctggctacca gcttcatgta cggcagcaga accttcatca accacaccca
     1681 gggcatcccc gacttcttta agcagtcctt ccctgagggc ttcacatggg agagagtcac
     1741 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
     1801 cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca
     1861 gaagaaaaca ctcggctggg aggccaacac cgagatgctg taccccgctg acggcggcct
     1921 ggaaggcaga agcgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt
     1981 caagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta
     2041 tgtggaccac agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca
     2101 cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca aacttaatgg
     2161 atccggagcc acgaacttct ctctgttaaa gcaagcagga gacgtggaag aaaaccccgg
     2221 tcctcacagc cagaagaggg tggtcgtgct gggctccgga gtcatcgggc tctctagcgc
     2281 cctgattctc gctagaaaag gctactccgt gcatatcctg gccagagact tgcctgagga
     2341 tgtcagctcc cagaccttcg caagcccctg ggccggagct aactggacac catttatgac
     2401 cctgacagac ggacccagac aagccaagtg ggaagagtcc accttcaaga aatgggtgga
     2461 gctggttccc acaggccacg ctatgtggct gaaagggacc agaaggtttg cccagaatga
     2521 ggatggcctc ctgggacatt ggtacaagga catcacccct aactataggc cacttccaag
     2581 ctctgaatgt ccacctggag ccattggagt gacatacgac accctgagcg tccacgctcc
     2641 caagtattgc caataccttg ccagagagct gcagaaactt ggggctacct tcgaaaggag
     2701 aacagtgact agcctggagc aagccttcga tggagctgac ctcgtggtca acgccaccgg
     2761 gctgggagct aagtccattg ccggcatcga tgaccaggct gccgagccca ttaggggaca
     2821 gaccgtgctt gtcaaatctc cctgtaagag gtgcacaatg gatagctccg accctgcttc
     2881 tcccgcttat atcattccta gaccaggagg agaagtgatc tgtggcggaa cctacggagt
     2941 cggcgattgg gacctgagcg tgaatcccga aacagtccaa aggattctca agcactgcct
     3001 gagacttgac cctaccatta gctccgatgg aactatcgag ggcattgaag tgctgagaca
     3061 taacgttggc ctggcacccg ccaggagagg aggacctagg gtggaggctg aaagaatcgt
     3121 cttgcccctg gaccggacca agagccctct ctctctggga agagggtccg cacgggctgc
     3181 aaaagagaag gaagtgaccc ttgtccacgc ctacggcttt agtagcgcag ggtatcagca
     3241 atcctgggga gccgctgagg acgtggctca gctggtcgat gaagccttcc aaagatacca
     3301 cggcgcagct agggagagcc tgccccccct ggagcggctg accctgtaaa agcttatcga
     3361 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3421 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3481 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3541 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3601 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3661 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3721 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3781 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3841 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3901 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3961 agcgctgctc gagagatctg tgatagcggc catcaagctg gccgcgactc tagatcataa
     4021 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4081 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4141 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4201 attctagttg tggtttgtcc aaactcatca atgtatcagc ttatcgatac cgcatgcacg
     4261 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4321 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4381 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4441 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4501 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4561 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4621 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4681 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4741 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4801 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     4861 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4921 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4981 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5041 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5101 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5161 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5221 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5281 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5341 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5401 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5461 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5521 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5581 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5641 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5701 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5761 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5821 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5881 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5941 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6001 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6061 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6121 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6181 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6241 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6301 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6361 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6421 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6481 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6541 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6601 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6661 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6721 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6781 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6841 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6901 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6961 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
