LOCUS       Exported                6479 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of mCerulean from CB7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pENN.AAV.CB7.CI.mCerulean.WPRE.RBG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6479)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6479)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6479
                     /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"
     regulatory      1954..1963
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1960..2679
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2005..2026)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2266..2285)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2613..2634
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2702..3290
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2755..2775)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3173..3184)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3316..3335)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3381..3436
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3529..3658
                     /label=AAV2 ITR
     primer_bind     complement(3677..3694)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3677..3693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3686..3708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3835..4290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3922..3941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4132..4153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4316..4420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4421..5281
                     /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(4639..4658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5452..6040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5941..5960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6194..6211
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6328..6349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6364..6394
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6402..6418
                     /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     6407..6429
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6426..6442
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6426..6442
                     /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 ttcacgcgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     1981 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2041 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2101 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg
     2161 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2221 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2281 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2341 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacgccatc
     2401 agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     2461 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2521 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     2581 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2641 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccatca agcttatcga
     2701 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2761 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2821 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2881 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2941 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3001 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3061 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3121 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3181 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3301 tcgatgagga tccgatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3361 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3421 ttttttgtgt ctctcactcg gaagcaattc gttgatctga atttcgacca cccataatac
     3481 ccattaccct ggtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     3541 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3601 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc
     3661 ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     3841 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3901 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3961 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4021 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4141 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4201 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4261 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4321 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4381 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4441 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4501 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4561 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4621 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4681 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4741 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4801 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4861 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4921 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4981 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5041 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5101 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5161 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5221 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5281 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5341 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5401 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5461 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5521 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5581 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5641 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5701 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5761 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5821 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5881 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5941 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6001 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6061 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6121 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6181 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6241 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6301 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6361 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6421 tcacacagga aacagctatg accatgatta cgccagattt aattaaggcc ttaattagg
//
