LOCUS       sequence-379358-        6857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6857)
  AUTHORS   Leopold AV, Thankachan S, Yang C, Gerashchenko D, Verkhusha VV
  TITLE     A general approach for engineering RTKs optically controlled with 
            far-red light.
  JOURNAL   Nat Methods. 2022 Jul;19(7):871-880. doi: 
            10.1038/s41592-022-01517-z. Epub 2022 Jun 9.
  PUBMED    35681062
REFERENCE   2  (bases 1 to 6857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-022-01517-z"; journalName: "Nat Methods"; date: 
            "2022-07"; volume: "19"; issue: "7"; pages: "871-880"
COMMENT     Sequence Label: AAV2/9-shMBVR-HO1-Fd-Fnr-CW3SL
FEATURES             Location/Qualifiers
     source          1..6857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        complement(234..448)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(294..313)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     CDS             898..972
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1003..1029
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1753..1809
                     /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"
     CDS             1810..1884
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1915..1944
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2245..2301
                     /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"
     CDS             2302..2376
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     complement(3740..3760)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4120..4260
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4120..4249
                     /label=AAV2 ITR
     rep_origin      4335..4790
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4422..4441)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4632..4653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4807..4826)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4926..4948
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4986..5004)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5072..5176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5177..5245,5246..6037)
                     /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(5395..5414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6208..6796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6697..6716
                     /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 gcggccgcag atgctgcgtc gacggtatcg ataagcttaa aaagccaaat
      181 gtaggagtca ataatctctt gaattattga ctcctacatt tggcgggaga tctgtggtct
      241 catacagaac ttataagatt cccaaatcca aagacatttc acgtttatgg tgatttccca
      301 gaacacatag cgacatgcaa atattgcagg gcgccactcc cctgtccctc acagccatct
      361 tcctgccagg gcgcacgcgc gctgggtgtt cccgcctagt gacactgggc ccgcgattcc
      421 ttggagcggg ttgatgacgt cagcgttcga attctcgaca ttgattattg actagtgcag
      481 atgctgcaca cttgtggact aagtttgttc gcatcccctt ctccaacccc ctcagtacat
      541 caccctgggg gaacagggtc cacttgctcc tgggcccaca cagtcctgca gtattgtgta
      601 tataaggcca gggcaaagag gagcaggttt taaagtgaaa ggcaggcagg tgttggggag
      661 gcagttaccg gggcaacggg aacagggcgt ttcggaggtg gttgccatgg ggacctggat
      721 gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct
      781 cgtcagtcaa gccggttctc cgtttgcact caggagcacg ggcaggcgag tggcccctag
      841 ttctgggggc agcgtgaaaa gacggtatcg ataagcttga ttcgagctag cgccaccatg
      901 tcagtcctga cacctctgct gctgcgagga ctgactggaa gcgccaggcg cctgcctgtg
      961 ccacgagcaa aaatccactc actgggcggg agcggaggca tgtaccccta tgatgtccct
     1021 gactacgcca tgactaccag cctggctaca aagctgaggg aaggcactaa gaaagctcat
     1081 accatggcag agaacgtggg gtttgtccgg tgtttcctga agggaactgt ggagaaaagc
     1141 tcctatagaa agctggtggc ctctctgtac cacgtctata gtgctatgga gcaggaaatg
     1201 gagcgcctga aagaccatcc aatcgtgggc aagatctact ttcccgaact gaacaggaag
     1261 tctagtctgg agcgcgatct gacatactat ttcgggtcca attggaggga ggaaattccc
     1321 ccttctcccg caactcaggc ctacgtggct cgcatccacg aagtcgcaaa caccgcccct
     1381 gagctgctgg tggcccatag ttacacacgg tatctgggag acctgtcagg gggacagatc
     1441 ctgaagggca ttgctgaacg ggcaatgaat ctgcaggatg gggagggaac cgccttctac
     1501 agattcgaaa gtatctcaga cgagaaggcc ttcaaacagc tgtatcgaca gcggctggat
     1561 gaactgcccg tggacgaggc tacagcagat agaattgtgg acgaggctaa cgctgcattt
     1621 ggcatgaata tgaagatctt ccaggaactg gagggcaacc tgatccgggc cattgggcag
     1681 ctgctgttta ataccctgac acgacggaaa cagagaggca gcaccgagct ggccacagct
     1741 gatggctcag gggcaaccaa cttcagcctg ctgaagcagg ccggagacgt ggaagagaat
     1801 ccaggaccta tgtccgtcct gaccccactg ctgctgaggg gactgacagg atctgcaaga
     1861 aggctgccag tgcccagagc caaaattcac tccctgggag gctctggagg catggagcag
     1921 aagctgatca gcgaggaaga tctgatggcc tcctatacag tgaaactgat cactcccgac
     1981 ggcgaatcaa gcattgagtg ctccgacgat acctacatcc tggatgcagc tgaggaagca
     2041 gggctggacc tgccttattc ttgtagggca ggagcctgca gtacttgtgc aggcaagatt
     2101 accgccggga gcgtggatca gagtgaccag tcatttctgg acgatgacca gatcgaagct
     2161 ggctacgtgc tgacatgcgt cgcatatccc acttccgatt gtactattga gacccacaaa
     2221 gaggaagacc tgtacggatc cggcgctaca aacttctctc tgctgaagca ggcaggggac
     2281 gtggaggaaa atcctggacc aatgagtgtc ctgactcccc tgctgctgag aggactgacc
     2341 ggaagcgccc gccgactgcc cgtgcctaga gctaagatcc atagtctggg gggatcaggc
     2401 gggatgtaca gcccaggata tgtggccacc agcagccggc agtccgatgc aggaaacagg
     2461 ctgttcgtgt acgaggtcat cggcctgagc cagtccacta tgaccgatgg gctggactat
     2521 cccattcgga gatctggaag tacattcatc actgtgcctc tgaaacgaat gaatcaggag
     2581 atgaggcgca tcacccggat gggaggcaaa atcgtgtcaa ttaagcctct ggaaggcgac
     2641 agcccactgc cacacacaga gggaattgcc aagccatcac agagcgaagg gtccggatct
     2701 gaggctgtgg caaaccctgc tccagaaagc aacaagacta tgacaactac ccctaaggag
     2761 aagaaagccg atgacatccc agtgaacatc tacaggccaa aaacccccta tattgggaag
     2821 gtgctggaaa attacccact ggtccgcgag ggagctatcg gaaccgtgca gcacctgaca
     2881 tttgatctgt ctgccggcga cctgcggtat ctggaagggc agagtatcgg aatcattcca
     2941 cccggagagg atgacaaggg caaacctcac aaactgaggc tgtacagcat tgcctccaca
     3001 cgccatgggg atttcggaga tgacaagact gtgagcctgt gcgtcagaca gctggaatac
     3061 cagaacgaag caggagagac cgtgcaggga gtctgctcca catacctgtg caatatcaaa
     3121 gagggcgatg acatcgccat taccggccca gtggggaagg aaatgctgct gcctccagat
     3181 gaggacgcaa acattgtcat gctggccaca ggcactggga tcgccccctt tcgggctttc
     3241 ctgtggagaa tgttcaaaga acagcacgag gactacaagt ttaaaggcct ggcctggctg
     3301 atcttcggga ttccaaagtc tgagaatatc ctgtataaag atgacctgga aaagatggca
     3361 gccgagtttc ccgataactt ccgcctgaca tacgctatta gccgagaaca gcagaatgca
     3421 gaggggggac gaatgtatat ccagcaccgg gtggccgaga acgctgagga actgtggaac
     3481 ctgatgcaga atccaaaaac ccatacatac atgtgcggac tgaagggcat ggaacccggg
     3541 atcgacgagg cctttaccgc cctggctgaa cagaatggaa aggagtggac aactttccag
     3601 agagagatga agaaagaaca tagatggcac gtcgagacct acatggcttc aatgacagga
     3661 ggacagcaga tggggtagct cgagataatc aacctctgga ttacaaaatt tgtgaaagat
     3721 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3781 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3841 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3901 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg
     3961 ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4021 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4081 gttcaggggg agatgtggga ggttttttaa agcggccgca ggaaccccta gtgatggagt
     4141 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4201 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4261 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4321 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4381 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4441 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4501 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4561 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4621 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4681 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4741 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4801 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4861 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4921 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4981 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5041 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5101 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5161 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5221 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5281 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5341 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5401 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5461 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5521 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5581 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5641 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5701 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5761 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5821 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5881 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5941 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6001 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6061 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6121 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6181 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6241 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6301 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6361 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6421 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6481 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6541 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6601 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6661 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6721 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6781 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6841 gccttttgct cacatgt
//
