LOCUS       Exported                6253 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 6253)
  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..6253
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        145..448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     146..172
                     /label=hCMVenh_f(138tg)
     primer_bind     351..365
                     /label=hCMVenh_f(138tg)
     promoter        449..652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     675..697
                     /label=RFP-F-NheI
     primer_bind     679..697
                     /label=mRuby2-NheI-Fw
     primer_bind     679..697
                     /label=Катя*mRuby2-NheI-Fw
     primer_bind     679..696
                     /label=RFP_F_EcoRI
     regulatory      679..688
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             685..1395
                     /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(1040..1067)
                     /label=TagBFP-seq-rev62
     primer_bind     1040..1067
                     /label=TagBFP-seq-dir62
     primer_bind     complement(1372..1401)
                     /label=RFP-R-BamHI
     primer_bind     complement(1372..1395)
                     /label=RFP_R_HindIII
     primer_bind     1396..1413
                     /label=p2A-BamHI-Fw
     primer_bind     1396..1413
                     /label=Катя*p2A-BamHI-Fw
     CDS             1405..1461
                     /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     1442..1485
                     /label=Юра*F_DAO_overlap
     primer_bind     complement(1442..1461)
                     /label=p2A-Rv
     primer_bind     complement(1442..1461)
                     /label=Катя*p2A-Rv
     CDS             1462..2556
                     /codon_start=1
                     /label=DAAOmut
                     /translation="HSQKRVVVLGSGVIGLSSALILARKGYSVHILARDLPEDVSSQTF
                     ASPWAGANWTPFMTLTDGPRQAKWEESTFKKWVELVPTGHAMWLKGTRRFAQNEDGLLG
                     HWYKDITPNYRPLPSSECPPGAIGVTYDTLSVHAPKYCQYLARELQKLGATFERRTVTS
                     LEQAFDGADLVVNATGLGAKSIAGIDDQAAEPIRGQTVLVKSPCKRCTMDSSDPASPAY
                     IIPRPGGEVICGGTYGVGDWDLSVNPETVQRILKHCLRLDPTISSDGTIEGIEVLRHNV
                     GLAPARRGGPRVEAERIVLPLDRTKSPLSLGRGSARAAKEKEVTLVHAYGFSSAGYQQS
                     WGAAEDVAQLVDEAFQRYHGAARES"
     primer_bind     1477..1494
                     /label=Юльчи и Фены, cat#180
     primer_bind     complement(1994..2010)
                     /label=DAO-MidR, Юльчи и Фены, cat#47
     primer_bind     1994..2010
                     /label=DAO-MidF, Юльчи и Фены, cat#46
     primer_bind     2095..2113
                     /label=DAO-MV-F, Юльчи и Фены, cat#182
     primer_bind     2301..2331
                     /label=Юльчи и Фены, cat#259
     primer_bind     complement(2311..2326)
                     /label=DAAOmut_R
     misc_feature    2311..2312
                     /label=mut
     primer_bind     complement(2496..2512)
                     
     primer_bind     complement(2531..2556)
                     /label=R_DAO_St_EcoRI
     primer_bind     complement(2540..2592)
                     /label=Юльчи и Фены, cat#101
     CDS             2557..2583
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2569..2586)
                     /label=Юльчи и Фены, cat#92
     misc_feature    2599..3187
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2602..2623)
                     /label=pLenti-R #136
     primer_bind     complement(2649..2668)
                     /label=pLu-Far-R #143
     primer_bind     complement(2652..2672)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(2664..2685)
                     /label=pLVT1-WPRE-Rv (Дина)
     CDS             complement(3070..3081)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3356..3437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3393..3412)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3447..3466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3516..3656
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3516..3645
                     /label=AAV2 ITR
     rep_origin      3731..4186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3818..3837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4028..4049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4203..4222)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4322..4344
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4382..4400)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4468..4572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4573..5433
                     /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(4791..4810)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5604..6192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6054..6063)
                     /label=DAO-MV-F, Юльчи и Фены, cat#182
     primer_bind     6093..6112
                     /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 gcgtcgttac ataacttacg gtaaatggcc cgcctggctg
      181 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      301 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      361 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      421 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      481 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      541 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      601 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      661 gaaccgtcag atccgctagc caccatggtg tctaagggcg aagagctgat taaggagaac
      721 atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc
      781 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
      841 ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcagaacc
      901 ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc tgagggcttc
      961 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
     1021 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc
     1081 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccaacaccga gatgctgtac
     1141 cccgctgacg gcggcctgga aggcagaagc gacatggccc tgaagctcgt gggcgggggc
     1201 cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa gaacctcaag
     1261 atgcccggcg tctactatgt ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag
     1321 acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg
     1381 gggcacaaac ttaatggatc cggagccacg aacttctctc tgttaaagca agcaggagac
     1441 gtggaagaaa accccggtcc tcacagccag aagagggtgg tcgtgctggg ctccggagtc
     1501 atcgggctct ctagcgccct gattctcgct agaaaaggct actccgtgca tatcctggcc
     1561 agagacttgc ctgaggatgt cagctcccag accttcgcaa gcccctgggc cggagctaac
     1621 tggacaccat ttatgaccct gacagacgga cccagacaag ccaagtggga agagtccacc
     1681 ttcaagaaat gggtggagct ggttcccaca ggccacgcta tgtggctgaa agggaccaga
     1741 aggtttgccc agaatgagga tggcctcctg ggacattggt acaaggacat cacccctaac
     1801 tataggccac ttccaagctc tgaatgtcca cctggagcca ttggagtgac atacgacacc
     1861 ctgagcgtcc acgctcccaa gtattgccaa taccttgcca gagagctgca gaaacttggg
     1921 gctaccttcg aaaggagaac agtgactagc ctggagcaag ccttcgatgg agctgacctc
     1981 gtggtcaacg ccaccgggct gggagctaag tccattgccg gcatcgatga ccaggctgcc
     2041 gagcccatta ggggacagac cgtgcttgtc aaatctccct gtaagaggtg cacaatggat
     2101 agctccgacc ctgcttctcc cgcttatatc attcctagac caggaggaga agtgatctgt
     2161 ggcggaacct acggagtcgg cgattgggac ctgagcgtga atcccgaaac agtccaaagg
     2221 attctcaagc actgcctgag acttgaccct accattagct ccgatggaac tatcgagggc
     2281 attgaagtgc tgagacataa cgttggcctg gcacccgcca ggagaggagg acctagggtg
     2341 gaggctgaaa gaatcgtctt gcccctggac cggaccaaga gccctctctc tctgggaaga
     2401 gggtccgcac gggctgcaaa agagaaggaa gtgacccttg tccacgccta cggctttagt
     2461 agcgcagggt atcagcaatc ctggggagcc gctgaggacg tggctcagct ggtcgatgaa
     2521 gccttccaaa gataccacgg cgcagctagg gagagcctgc cccccctgga gcggctgacc
     2581 ctgtaaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2641 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2701 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2761 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2821 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2881 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2941 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3001 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     3061 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3121 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3181 tccccgcatc gataccgagc gctgctcgag agatctgtga tagcggccat caagctggcc
     3241 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3301 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3361 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3421 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcagctta
     3481 tcgataccgc atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     3541 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3601 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3661 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3721 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3781 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     3841 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3901 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     3961 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4021 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4081 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4141 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4201 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4261 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4321 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4381 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4441 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4501 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4561 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4621 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4681 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4741 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4801 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4861 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4921 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4981 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5041 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5101 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5161 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5221 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5281 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5341 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5401 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5461 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5521 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5581 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5641 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5701 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5761 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5821 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5881 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5941 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6001 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6061 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6121 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6181 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6241 tttgctcaca tgt
//
