LOCUS       CDS-G002_pAAV-CA        6237 bp DNA     circular SYN 26-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6237)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Oct 26, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6237
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     primer_bind     complement(270..281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     331..342
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        701..978
                     /label=chicken beta-actin promoter
     primer_bind     1258..1280
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1342..1361
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1368..1427
                     /label=Flag-miniSOG-F2
     CDS             1400..1426
                     /codon_start=1
                     /label=FLAG
                     /translation="MDYKDDDDK"
     primer_bind     1407..1466
                     /label=Flag-miniSOG-F1
     CDS             1442..1759
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     primer_bind     complement(1767..1826)
                     /label=miniSOG-C-Far-R1
     CDS             1784..1840
                     /codon_start=1
                     /label=C-Far
                     /translation="KLNPPDESGPGCMSCCVLS"
     misc_feature    1784..1820
                     /label=Cytoplasmic
     primer_bind     complement(1813..1865)
                     /label=Flag-miniSOG-C-Far-R2
     CDS             1847..1900
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    1916..2620
                     /label=dTomato
     misc_feature    2631..3219
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2684..2704)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3102..3113)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3262..3383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3273..3292)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3327..3346
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3545..3674
                     /label=AAV2 ITR
     primer_bind     complement(3591..3602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3652..3663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3693..3710)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3693..3709)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3702..3724)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3851..4306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3938..3957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4148..4169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4332..4436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4437..5297
                     /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(4655..4674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5468..6056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5957..5976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6210..6227
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatcgta ccattgacgt caataatgac gtatgttccc
      481 atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt acggtaaact
      541 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      601 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      661 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      721 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      781 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      841 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      901 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      961 cgaagcgcgc ggcgggcggg agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc
     1021 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     1081 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     1141 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
     1201 cggctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1261 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1321 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1381 aagaattgga tccgccacca tggattacaa ggatgacgac gataagggcg gaggtggttc
     1441 tatggagaaa agtttcgtga taactgatcc acggctgcca gacaatccca tcatcttcgc
     1501 atccgatggc ttcctggagc tgaccgagta ttccagagag gagatcctgg gccgcaatgg
     1561 ccgctttctg cagggaccag agacagacca ggccacagtg cagaagattc gcgatgccat
     1621 tagagatcag cgcgagatta ccgtgcagct gataaactac acaaaaagcg ggaagaaatt
     1681 ctggaacctc ctgcacctcc agcccatgag ggaccagaag ggtgagctcc agtatttcat
     1741 cggagtgcag ctggatggat caggaggagg ctcaggagga ggaaagctga accctcctga
     1801 tgagagtggc cccggctgca tgagctgctg tgtgctctcc agatctgagg gtcggggctc
     1861 tctgctcaca tgtggcgacg tcgaggagaa tcccggaccg gccccggggt cgacaatggt
     1921 gtctaagggt gaggaagtaa ttaaggagtt tatgagattc aaagttagaa tggagggtag
     1981 catgaacggg cacgagtttg aaattgaagg tgagggtgaa ggaagaccat atgagggcac
     2041 acaaaccgcc aagctcaagg ttaccaaagg tggccccctt cctttcgcct gggacatcct
     2101 cagccctcaa ttcatgtacg gtagcaaggc ttacgtcaaa caccctgctg atatacctga
     2161 ttacaaaaaa ctgtcatttc ctgaaggatt taagtgggaa cgagtaatga attttgagga
     2221 tggtgggctg gtcactgtca cgcaagactc ctctctgcag gacggcaccc tgatttacaa
     2281 ggtgaagatg cgcggaacaa attttccccc cgatggcccc gtgatgcaga agaagaccat
     2341 ggggtgggag gcctctacag aacgactgta cccccgcgat ggcgttctta aaggcgaaat
     2401 ccatcaggcg cttaaactca aagatggcgg gcactacctg gttgagttca aaactatata
     2461 tatggccaaa aagccggttc agctgcctgg ctactactac gtggacacca agttggacat
     2521 cacgagtcac aacgaggact acacaatcgt cgagcagtac gaacgctcag aaggtcggca
     2581 tcacctgttc ttgtatggca tggatgagct gtataagtaa gcttatcgat aatcaacctc
     2641 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2701 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2761 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2821 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2881 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2941 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3001 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     3061 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3121 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3181 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacccgggc
     3241 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3301 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3361 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3421 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     3481 ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     3541 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3601 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3661 gagcgagcgc gcagccttaa ttaacctaat tcactggccg tcgttttaca acgtcgtgac
     3721 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     3781 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3841 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3901 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3961 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4021 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4081 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4141 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4201 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4261 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     4321 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4381 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4441 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4501 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4561 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4621 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4681 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4741 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4801 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4861 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4921 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4981 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5041 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5101 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5161 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5221 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5281 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5341 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5401 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5461 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5521 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5581 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5641 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5701 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5761 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5821 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5881 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5941 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6001 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6061 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6121 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6181 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
