LOCUS       MB46pscAAV-CAG-G        5077 bp    DNA     circular SYN 28-JAN-2022
DEFINITION  recombinant AAV vector packaging self-complementary GFP under the 
            CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5077)
  AUTHORS   Kay MA
  TITLE     Kay lab - unpublished AAV vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5077)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 26, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pscAAV-CAG-GFP
FEATURES             Location/Qualifiers
     source          1..5077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..161
                     /label=ITR is about here
                     /note="color: #a6acb3"
     primer_bind     complement(46..57)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     85..96
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     120..143
                     /label=ITR_W3SL_gib_r
                     /note="color: black; sequence: 
                     caactccatcactaggggttcctgcggcccaaggtaccttgtttaaaaaacctcccaca
                     tctccccc; added: 2020-11-06"
     misc_feature    142..161
                     /label=giboson overhang
                     /note="color: #a6acb3"
     promoter        complement(164..182)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    183..232
                     /label=stuffer
                     /note="color: #a6acb3"
     promoter        233..251
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(260..284)
                     /label=rBC6-P11
                     /note="color: #0000ff; direction: LEFT"
     primer_bind     complement(287..311)
                     /label=rBC6-P12
                     /note="color: #0000ff; direction: LEFT"
     primer_bind     complement(314..338)
                     /label=rBC6-P13
                     /note="color: #0000ff; direction: LEFT"
     primer_bind     complement(341..365)
                     /label=rBC6-P14
                     /note="color: #0000ff; direction: LEFT"
     misc_feature    368..390
                     /label=gibson overhang
                     /note="color: #a6acb3"
     misc_feature    369..618
                     /label=truncated CMV enhancer
                     /note="color: #ccffff"
     primer_bind     complement(396..415)
                     /label=T7BConlyKay_1
                     /note="color: black; sequence: CCTATTGGCGTTACTATGGG; added:
                     2021-06-01"
     promoter        619..896
                     /label=chicken beta-actin promoter
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(620..639)
                     /label=MB_seq_24
                     /note="color: black; sequence: agaacgtggggctcacctcg; added:
                     2021-06-11"
     primer_bind     648..662
                     /label=ITR_W3SL_gib_r
                     /note="color: black; sequence: 
                     caactccatcactaggggttcctgcggcccaaggtaccttgtttaaaaaacctcccaca
                     tctccccc; added: 2020-11-06"
     primer_bind     1175..1197
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
                     /note="color: black; sequence: GGTTCGGCTTCTGGCGTGTGACC"
     primer_bind     1259..1278
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
                     /note="color: black; sequence: GCAACGTGCTGGTTATTGTG"
     regulatory      1318..1327
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             join(1324..1326,1327..1329,1330..2043)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1324 .. 1326 / #05fd14
                        2: 1327 .. 1329 / #05fd14 / 1a
                        3: 1330 .. 2043 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1369..1390)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(1630..1649)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     1977..1998
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     primer_bind     2065..2085
                     /label=T7BConlyKay_2
                     /note="color: black; sequence: GCGATCTCGATGCTTTATTTG; 
                     added: 2021-06-01"
     primer_bind     2087..2106
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     2094..2114
                     /label=MB_seq_43
                     /note="color: black; sequence: gtgatgctattgctttatttg; 
                     added: 2021-06-11"
     primer_bind     complement(2182..2207)
                     /label=ITR_W3SL_gib_r
                     /note="color: black; sequence: 
                     caactccatcactaggggttcctgcggcccaaggtaccttgtttaaaaaacctcccaca
                     tctccccc; added: 2020-11-06"
     misc_feature    2183..2207
                     /label=gibson overhang
                     /note="color: #a6acb3"
     misc_feature    2208..2264
                     /label=stuffer
                     /note="color: #a6acb3"
     promoter        2265..2283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2284..2421
                     /label=ITR is about here
                     /note="color: #a6acb3"
     misc_feature    2288..2307
                     /label=gibson overhang
                     /note="color: #a6acb3"
     primer_bind     complement(2311..2322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     2372..2383
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     primer_bind     complement(2432..2449)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(2432..2448)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(2441..2463)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     primer_bind     complement(2657..2676)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /note="color: black; sequence: CGGCATCAGAGCAGATTGTA"
     primer_bind     2776..2798
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
                     /note="color: black; sequence: CCGGGAGCTGCATGTGTCAGAGG"
     primer_bind     complement(2836..2854)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
                     /note="color: black; sequence: AATAGGCGTATCACGAGGC"
     promoter        2922..3026
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(3027..3095,3096..3887)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3027 .. 3095 / #ccffcc / signal sequence
                        2: 3096 .. 3887 / #ccffcc
                     Cleavage site after base 3095"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3245..3264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      4058..4646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     4547..4566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     4800..4817
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     protein_bind    4934..4955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        join(4970..4975,4976..4993,4994..5000)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 4970 .. 4975 / #ffffff / -35
                        2: 4976 .. 4993 / #ffffff
                        3: 4994 .. 5000 / #ffffff / -10"
     protein_bind    5008..5024
                     /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)."
                     /note="color: #31849b"
     primer_bind     5013..5035
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     5032..5048
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     5032..5048
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgggcgacca aaggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctggaggggt ggagtcgtga cctcctatag tgagtcgtat
      181 tacccggcca ggaaccgtaa aaaggacttg cccgcgttgc tggcgtttaa cgatttaggt
      241 gacactatag aatacacggg tggtaaccgt attcattgga tttatgtgta tttgccagtg
      301 atggcaaatt gccaaggctg ctacggacga tgtggaagat ggcccaagag catctggaga
      361 gacaaaggcc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      661 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      721 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
      781 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      841 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      901 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      961 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     1021 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
     1081 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
     1141 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1201 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1261 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc accggtcgcc
     1321 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1381 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1441 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1501 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1561 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1621 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1681 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1741 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1801 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1861 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1921 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1981 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2041 taaagcggcc gctaggcctc acctgcgatc tcgatgcttt atttgtgaaa tttgtgatgc
     2101 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2161 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaatca ctgacgttaa
     2221 acgccagcaa cgcgggcaag tcctttttac ggttcctggc cgggtaatac gactcactat
     2281 aggctagtcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     2341 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg
     2401 acagatccgg gcccgcatgc gtcgacaatt cactggccgt cgttttacaa cgtcgtgact
     2461 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2521 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2581 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2641 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2701 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2761 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2821 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2881 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2941 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3001 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3061 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3121 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3181 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3241 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3301 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3361 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3421 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3481 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3541 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3601 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3661 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3721 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3781 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3841 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3901 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3961 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4021 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4081 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4141 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4201 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4261 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4321 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4381 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4441 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4501 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4561 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4621 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4681 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4741 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4801 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     4861 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     4921 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     4981 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5041 gctatgacca tgattacgcc aagctctcga gatctag
//
