LOCUS       Exported                7975 bp ds-DNA     circular SYN 10-JAN-2022
DEFINITION  Rescued infectious variant from pAAV2_RaPID library; mCherry 
            insertion in place of the deleted amino acid sequence 453-GTTTQSR.
ACCESSION   .
VERSION     .
KEYWORDS    pRC_RR_VP1_r1c3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7975)
  AUTHORS   Judd J, Wei F, Nguyen PQ, Tartaglia LJ, Agbandje-McKenna M, Silberg 
            JJ, Suh J
  TITLE     Random Insertion of mCherry Into VP3 Domain of Adeno-associated 
            Virus Yields Fluorescent Capsids With no Loss of Infectivity.
  JOURNAL   Mol Ther Nucleic Acids. 2012 Nov 13;1:e54. doi: 
            10.1038/mtna.2012.46.
  PUBMED    23629029
REFERENCE   2  (bases 1 to 7975)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 10, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7975
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     70..86
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     71..87
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(121..137)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(121..137)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3512..4219
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3657..3675)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3925..3944
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4898..4909
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(5188..5208)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5188..5206)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5227..5243)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5227..5243)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5240..5262)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5251..5267
                     /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(5275..5305)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5320..5341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5458..5475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5629..6217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5709..5728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6388..7248)
                     /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     7011..7030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7249..7353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7379..7834)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7516..7537)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7728..7747
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7961..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7975..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
       61 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
      121 gatccactag ttctagagtc ctgtattaga ggtcacgtga gtgttttgcg acattttgcg
      181 acaccatgtg gtcacgctgg gtatttaagc ccgagtgagc acgcagggtc tccattttga
      241 agcgggaggt ttgaacgcgc agccgccatg ccggggtttt acgagattgt gattaaggtc
      301 cccagcgacc ttgacgagca tctgcccggc atttctgaca gctttgtgaa ctgggtggcc
      361 gagaaggaat gggagttgcc gccagattct gacatggatc tgaatctgat tgagcaggca
      421 cccctgaccg tggccgagaa gctgcagcgc gactttctga cggaatggcg tcgtgtgagt
      481 aaggccccgg aggccctttt ctttgtgcaa tttgagaagg gagagagcta cttccacatg
      541 cacgtgctcg tggaaaccac cggggtgaaa tctatggttt tgggacgttt cctgagtcag
      601 attcgcgaaa aactgattca gagaatttac cgcgggatcg agccgacttt gccaaactgg
      661 ttcgcggtca caaagaccag aaatggtgcc ggaggcggga acaaggtggt ggatgagtgc
      721 tacatcccca attacttgct ccccaaaacc cagcctgagc tccagtgggc gtggactaat
      781 atggaacagt atttaagcgc ctgtttgaat ctcacggagc gtaaacggtt ggtggcgcag
      841 catctgacgc acgtgtcgca gacgcaggag cagaacaaag agaatcagaa tcccaattct
      901 gatgcgccgg tgatcagatc aaaaacttca gccaggtaca tggagctggt cgggtggctc
      961 gtggacaagg ggattacctc ggagaagcag tggatacagg aggaccaggc ctcatacatc
     1021 tccttcaatg cggcctccaa ctcgcggtcc caaatcaagg ctgccttgga caatgcggga
     1081 aagattatga gcctgactaa aaccgccccc gactacctgg tgggccagca gcccgtggag
     1141 gacatttcca gcaatcggat ttataaaatt ttggaactaa acgggtacga tccccaatat
     1201 gcggcttccg tctttctggg atgggccacg aaaaagttcg gcaagaggaa caccatctgg
     1261 ctgtttgggc ctgcaactac cgggaagacc aacatcgcgg aggccatagc ccacactgtg
     1321 cccttctacg ggtgcgtaaa ctggaccaat gagaactttc ccttcaacga ctgtgtcgac
     1381 aagatggtga tctggtggga ggaggggaag atgaccgcca aggtcgtgga gtcggccaaa
     1441 gccattctcg gaggaagcaa ggtgcgcgtg gaccagaaat gcaagtcctc ggcccagata
     1501 gacccgactc ccgtgatcgt cacctccaac accaacatgt gcgccgtgat tgacgggaac
     1561 tcaacgacct tcgaacacca gcagccgttg caagaccgga tgttcaaatt tgaactcacc
     1621 cgccgtctgg atcatgactt tgggaaggtc accaagcagg aagtcaaaga ctttttccgg
     1681 tgggcaaagg atcacgtggt tgaggtggag catgaattct acgtcaaaaa gggtggagcc
     1741 aagaaaagac ccgcccccag tgacgcagat ataagtgagc ccaaacgggt gcgcgagtca
     1801 gttgcgcagc catcgacgtc agacgcggaa gcttcgatca actacgcaga caggtaccaa
     1861 aacaaatgtt ctcgtcacgt gggcatgaat ctgatgctgt ttccctgcag acaatgcgag
     1921 agaatgaatc agaattcaaa tatctgcttc actcacggac agaaagactg tttagagtgc
     1981 tttcccgtgt cagaatctca acccgtttct gtcgtcaaaa aggcgtatca gaaactgtgc
     2041 tacattcatc atatcatggg aaaggtgcca gacgcttgca ctgcctgcga tctggtcaat
     2101 gtggatttgg atgactgcat ctttgaacaa taaatgattt aaatcaggta tggctgccga
     2161 tggttatcta cccgattggc tcgaggacac tctctctgaa ggaataagac agtggtggaa
     2221 gctcaaacct ggcccaccac caccaaagcc cgcagagcgg cataaggacg acagcagggg
     2281 tcttgtgctt cctgggtaca agtacctcgg acccttcaac ggactcgaca agggagagcc
     2341 ggtcaacgag gcagacgccg cggccctcga gcacgacaaa gcctacgacc ggcagctcga
     2401 cagcggagac aacccgtacc tcaagtacaa ccacgccgac gcggagtttc aggagcgcct
     2461 taaagaagat acgtcttttg ggggcaacct cggacgagca gtcttccagg cgaaaaagag
     2521 ggttcttgaa cctctgggcc tggttgagga acctgttaag acggctccgg gaaaaaagag
     2581 gccggtagag cactctcctg tggagccaga ctcctcctcg ggaaccggaa aggcgggcca
     2641 gcagcctgca agaaaaagat tgaattttgg tcagactgga gacgcagact cagtacctga
     2701 cccccagcct ctcggacagc caccagcagc cccctctggt ctgggaacta ataccatggc
     2761 tacaggcagt ggcgcaccaa tggcagacaa taacgagggt gccgacggag tgggtaattc
     2821 ctcgggaaat tggcattgcg attccacatg gatgggcgac agagtcatca ccaccagcac
     2881 ccgaacctgg gccctgccca cctacaacaa ccacctctac aaacaaattt ccagccaatc
     2941 aggagcctcg aacgacaatc actactttgg ctacagcacc ccttgggggt attttgactt
     3001 caacagattc cactgccact tttcaccacg tgactggcaa agactcatca acaacaactg
     3061 gggattccga cccaagagac tcaacttcaa gctctttaac attcaagtca aagaggtcac
     3121 gcagaatgac ggtacgacga cgattgccaa taaccttacc agcacggttc aggtgtttac
     3181 tgactcggag taccagctcc cgtacgtcct cggctcggcg catcaaggat gcctcccgcc
     3241 gttcccagca gacgtcttca tggtgccaca gtatggatac ctcaccctga acaacgggag
     3301 tcaggcagta ggacgctctt cattttactg cctggagtac tttccttctc agatgctgcg
     3361 taccggaaac aactttacct tcagctacac ttttgaggac gttcctttcc acagcagcta
     3421 cgctcacagc cagagtctgg accgtctcat gaatcctctc atcgaccagt acctgtatta
     3481 cttgagcaga acaaacactc caagtgccgg catggtgagc aagggcgagg aggataacat
     3541 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     3601 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     3661 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     3721 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     3781 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     3841 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     3901 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagactatgg gctgggaggc
     3961 ctcctccgag cggatgtacc ccgaggacgg tgccctgaag ggcgagatca agcagaggct
     4021 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     4081 gcccgtgcag ctgcccggcg cttacaacgt caacatcaag ttggacatca cctcccacaa
     4141 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     4201 catggacgag ctgtacaagg gcgcccttca gttttctcag gccggagcga gtgacattcg
     4261 ggaccagtct aggaactggc ttcctggacc ctgttaccgc cagcagcgag tatcaaagac
     4321 atctgcggat aacaacaaca gtgaatactc gtggactgga gctaccaagt accacctcaa
     4381 tggcagagac tctctggtga atccgggccc ggctatggca agccacaagg acgatgaaga
     4441 aaagtttttt cctcagagcg gggttctcat ctttgggaag caaggctcag agaaaacaaa
     4501 tgtggacatt gaaaaggtca tgattacaga cgaagaggaa atcaggacaa ccaatcccgt
     4561 ggctacggag cagtatggtt ctgtatctac caacctccag agaggcaaca gacaagcagc
     4621 taccgcagat gtcaacacac aaggcgttct tccaggcatg gtctggcagg acagagatgt
     4681 gtaccttcag gggcccatct gggcaaagat tccacacacg gacggacatt ttcacccctc
     4741 tcccctcatg ggtggattcg gacttaaaca ccctcctcca cagattctca tcaagaacac
     4801 cccggtgcct gcgaatcctt cgaccacctt cagtgcggca aagtttgctt ccttcatcac
     4861 acagtactcc acgggacagg tcagcgtgga gatcgagtgg gagctgcaga aggaaaacag
     4921 caaacgctgg aatcccgaaa ttcagtacac ttccaactac aacaagtctg ttaatgtgga
     4981 ctttactgtg gacactaatg gcgtgtattc agagcctcgc cccattggca ccagatacct
     5041 gactcgtaat ctgtaattgg caggtcttgt taatcaataa accgtttaat tcgtttcagt
     5101 tgaactttgg tctctgcgta tttctttctt atctagtttc catgctctag agcggccgcc
     5161 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     5221 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5281 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5341 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     7381 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     7441 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     7501 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     7561 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     7621 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     7681 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     7741 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     7801 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     7861 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7921 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgtt
//
