LOCUS       sequence-135909-        7582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Adeno-associated virus VP2 with PIF6 insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7582)
  AUTHORS   Gomez EJ, Gerhardt K, Judd J, Tabor JJ, Suh J
  TITLE     Light-Activated Nuclear Translocation of Adeno-Associated Virus 
            Nanoparticles Using Phytochrome B for Enhanced, Tunable, and 
            Spatially Programmable Gene Delivery.
  JOURNAL   ACS Nano. 2015 Nov 30.
  PUBMED    26618393
REFERENCE   2  (bases 1 to 7582)
  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"; journalName: "ACS Nano. 
            2015 Nov 30."
COMMENT     Sequence Label: pVP2A-PIF6
FEATURES             Location/Qualifiers
     source          1..7582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(720..736)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(720..736)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5110..5121
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(5394..5414)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5394..5412)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5433..5449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5433..5449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5446..5468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5457..5473
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5481..5487,5488..5505,5506..5511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5526..5547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5664..5681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5835..6423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5915..5934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6594..7385,7386..7454))
                     /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     7217..7236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7455..7559)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
      721 atccactagt tctagagtcc tgtattagag gtcacgtgag tgttttgcga cattttgcga
      781 caccatgtgg tcacgctggg tatttaagcc cgagtgagca cgcagggtct ccattttgaa
      841 gcgggaggtt tgaacgcgca gccgccatgc cggggtttta cgagattgtg attaaggtcc
      901 ccagcgacct tgacgagcat ctgcccggca tttctgacag ctttgtgaac tgggtggccg
      961 agaaggaatg ggagttgccg ccagattctg acatggatct gaatctgatt gagcaggcac
     1021 ccctgaccgt ggccgagaag ctgcagcgcg actttctgac ggaatggcgc cgtgtgagta
     1081 aggccccgga ggcccttttc tttgtgcaat ttgagaaggg agagagctac ttccacatgc
     1141 acgtgctcgt ggaaaccacc ggggtgaaat ccatggtttt gggacgtttc ctgagtcaga
     1201 ttcgcgaaaa actgattcag agaatttacc gcgggatcga gccgactttg ccaaactggt
     1261 tcgcggtcac aaagaccaga aatggcgccg gaggcgggaa caaggtggtg gatgagtgct
     1321 acatccccaa ttacttgctc cccaaaaccc agcctgagct ccagtgggcg tggactaata
     1381 tggaacagta tttaagcgcc tgtttgaatc tcacggagcg taaacggttg gtggcgcagc
     1441 atctgacgca cgtgtcgcag acgcaggagc agaacaaaga gaatcagaat cccaattctg
     1501 atgcgccggt gatcagatca aaaacttcag ccaggtacat ggagctggtc gggtggctcg
     1561 tggacaaggg gattacctcg gagaagcagt ggatccagga ggaccaggcc tcatacatct
     1621 ccttcaatgc ggcctccaac tcgcggtccc aaatcaaggc tgccttggac aatgcgggaa
     1681 agattatgag cctgactaaa accgcccccg actacctggt gggccagcag cccgtggagg
     1741 acatttccag caatcggatt tataaaattt tggaactaaa cgggtacgat ccccaatatg
     1801 cggcttccgt ctttctggga tgggccacga aaaagttcgg caagaggaac accatctggc
     1861 tgtttgggcc tgcaactacc gggaagacca acatcgcgga ggccatagcc cacactgtgc
     1921 ccttctacgg gtgcgtaaac tggaccaatg agaactttcc cttcaacgac tgtgtcgaca
     1981 agatggtgat ctggtgggag gaggggaaga tgaccgccaa ggtcgtggag tcggccaaag
     2041 ccattctcgg aggaagcaag gtgcgcgtgg accagaaatg caagtcctcg gcccagatag
     2101 acccgactcc cgtgatcgtc acctccaaca ccaacatgtg cgccgtgatt gacgggaact
     2161 caacgacctt cgaacaccag cagccgttgc aagaccggat gttcaaattt gaactcaccc
     2221 gccgtctgga tcatgacttt gggaaggtca ccaagcagga agtcaaagac tttttccggt
     2281 gggcaaagga tcacgtggtt gaggtggagc atgaattcta cgtcaaaaag ggtggagcca
     2341 agaaaagacc cgcccccagt gacgcagata taagtgagcc caaacgggtg cgcgagtcag
     2401 ttgcgcagcc atcgacgtca gacgcggaag cttcgatcaa ctacgcagac aggtaccaaa
     2461 acaaatgttc tcgtcacgtg ggcatgaatc tgatgctgtt tccctgcaga caatgcgaga
     2521 gaatgaatca gaattcaaat atctgcttca ctcacggaca gaaagactgt ttagagtgct
     2581 ttcccgtgtc agaatctcaa cccgtttctg tcgtcaaaaa ggcgtatcag aaactgtgct
     2641 acattcatca tatcatggga aaggtgccag acgcttgcac tgcctgcgat ctggtcaatg
     2701 tggatttgga tgactgcatc tttgaacaat aaatgattta aatcaggtct ggctgccgat
     2761 ggttatcttc cagattggct cgaggacact ctctctgaag gaataagaca gtggtggaag
     2821 ctcaaacctg gcccaccacc accaaagccc gcagagcggc ataaggacga cagcaggggt
     2881 cttgtgcttc ctgggtacaa gtacctcgga cccttcaacg gactcgacaa gggagagccg
     2941 gtcaacgagg cagacgccgc ggccctcgag cacgacaaag cctacgaccg gcagctcgac
     3001 agcggagaca acccgtacct caagtacaac cacgccgacg cggagtttca ggagcgcctt
     3061 aaagaagata cgtcttttgg gggcaacctc ggacgagcag tcttccaggc gaaaaagagg
     3121 gttcttgaac ctctgggcct ggttgaggaa cctgttaaga tgcggccgat gatgttcctt
     3181 cctactgatt attgttgcag actgagcgac caggaataca tggaactcgt cttcgagaac
     3241 ggacagatac tcgcaaaagg ccagaggtca aatgttagtc tccataatca gcggacgaaa
     3301 agcatcatgg atctgtatga ggccgaatac aacgaagatt ttatgaaaag tattatccat
     3361 ggagggggtg gcgctattac caacctggga gatacccaag tggtcccaca gtcccacgta
     3421 gcagccgctc acgagaccaa tatgctggag tccaacaaac acgtagacac gcgtgctccg
     3481 ggaaaaaaga ggccggtaga gcactctcct gtggagccag actcctcctc gggaaccgga
     3541 aaggcgggcc agcagcctgc aagaaaaaga ttgaattttg gtcagactgg agacgcagac
     3601 tcagtacctg acccccagcc tctcggacag ccaccagcag ccccctctgg tctgggaact
     3661 aatacgctgg ctacaggcag tggcgcacca ctggcagaca ataacgaggg cgccgacgga
     3721 gtgggtaatt cctcgggaaa ttggcattgc gattccacat ggctgggcga cagagtcatc
     3781 accaccagca cccgaacctg ggccctgccc acctacaaca accacctcta caaacaaatt
     3841 tccagccaat caggagcctc gaacgacaat cactactttg gctacagcac cccttggggg
     3901 tattttgact tcaacagatt ccactgccac ttttcaccac gtgactggca aagactcatc
     3961 aacaacaact ggggattccg acccaagaga ctcaacttca agctctttaa cattcaagtc
     4021 aaagaggtca cgcagaatga cggtacgacg acgattgcca ataaccttac cagcacggtt
     4081 caggtgttta ctgactcgga gtaccagctc ccgtacgtcc tcggctcggc gcatcaagga
     4141 tgcctcccgc cgttcccagc agacgtcttc atggtgccac agtatggata cctcaccctg
     4201 aacaacggga gtcaggcagt aggacgctct tcattttact gcctggagta ctttccttct
     4261 cagatgctgc gtaccggaaa caactttacc ttcagctaca cttttgagga cgttcctttc
     4321 cacagcagct acgctcacag ccagagtctg gaccgtctca tgaatcctct catcgaccag
     4381 tacctgtatt acttgagcag aacaaacact ccaagtggaa ccaccacgca gtcaaggctt
     4441 cagttttctc aggccggagc gagtgacatt cgggaccagt ctaggaactg gcttcctgga
     4501 ccctgttacc gccagcagcg agtatcaaag acatctgcgg ataacaacaa cagtgaatac
     4561 tcgtggactg gagctaccaa gtaccacctc aatggcagag actctctggt gaatccgggc
     4621 ccggccatgg caagccacaa ggacgatgaa gaaaagtttt ttcctcagag cggggttctc
     4681 atctttggga agcaaggctc agagaaaaca aatgtggaca ttgaaaaggt catgattaca
     4741 gacgaagagg aaatcaggac aaccaatccc gtggctacgg agcagtatgg ttctgtatct
     4801 accaacctcc agagaggcaa cagacaagca gctaccgcag atgtcaacac acaaggcgtt
     4861 cttccaggca tggtctggca ggacagagat gtgtaccttc aggggcccat ctgggcaaag
     4921 attccacaca cggacggaca ttttcacccc tctcccctca tgggtggatt cggacttaaa
     4981 caccctcctc cacagattct catcaagaac accccggtac ctgcgaatcc ttcgaccacc
     5041 ttcagtgcgg caaagtttgc ttccttcatc acacagtact ccacgggaca ggtcagcgtg
     5101 gagatcgagt gggagctgca gaaggaaaac agcaaacgct ggaatcccga aattcagtac
     5161 acttccaact acaacaagtc tgttaatgtg gactttactg tggacactaa tggcgtgtat
     5221 tcagagcctc gccccattgg caccagatac ctgactcgta atctgtaatt gcttgttaat
     5281 caataaaccg tttaattcgt ttcagttgaa ctttggtctc tgcgtatttc tttcttatct
     5341 agtttccatg ctctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     5401 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5461 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5521 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5581 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5641 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5701 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6241 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6481 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6541 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6601 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6661 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6721 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6781 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6841 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6901 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6961 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7021 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7081 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7141 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7201 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7261 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7321 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7381 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7441 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7501 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7561 acatttcccc gaaaagtgcc ac
//
