LOCUS       Exported                6502 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of mCherry from CB7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pENN.AAV.CB7.CI.mCherry.WPRE.RBG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6502)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6502)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6502
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        204..583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        585..862
                     /label=chicken beta-actin promoter
     intron          863..1879
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1803..1825
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1887..1906
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1962..2672
                     /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(2107..2125)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2375..2394
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2725..3313
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2778..2798)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3196..3207)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3339..3358)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3404..3459
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3552..3681
                     /label=AAV2 ITR
     primer_bind     complement(3700..3717)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3700..3716)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3709..3731)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3858..4313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3945..3964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4155..4176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4339..4443
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4444..5304
                     /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(4662..4681)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5475..6063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5964..5983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6217..6234
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6351..6372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6387..6417
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6425..6441
                     /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     6430..6452
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6449..6465
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6449..6465
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gtcgacattg attattgact agttattaat agtaatcaat
      241 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      301 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      361 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      421 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      481 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      541 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
      601 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
      661 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
      721 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
      781 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
      841 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
      901 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      961 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
     1021 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
     1081 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
     1141 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
     1201 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
     1261 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
     1321 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
     1381 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
     1441 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
     1501 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
     1561 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
     1621 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
     1681 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
     1741 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
     1801 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     1861 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     1921 tggcaaagaa ttcacgctag atttaagaag gagatataca tatggtgagc aagggcgagg
     1981 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     2041 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2101 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     2161 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     2221 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     2281 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     2341 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     2401 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     2461 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     2521 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     2581 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     2641 ccaccggcgg catggacgag ctgtacaagt aagcatgcaa gcttcgagga cggggtgaac
     2701 tacgcctgaa tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2761 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2821 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2881 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2941 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3001 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3061 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3121 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3181 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3241 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3301 gccgcctccc cgcatcgata ccgtcgatga ggatccgatc tttttccctc tgccaaaaat
     3361 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     3421 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaagcaa ttcgttgatc
     3481 tgaatttcga ccacccataa tacccattac cctggtagat aagtagcatg gcgggttaat
     3541 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3601 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3661 agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg
     3721 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3781 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3841 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3901 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3961 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4021 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4081 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4141 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4201 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4261 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4621 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4681 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4981 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5581 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6061 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6241 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6301 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6361 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6421 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga
     6481 tttaattaag gccttaatta gg
//
